Anti-cub domain-containing protein 1 (CDCP1) antibodies, antibody drug conjugates, and methods of use thereof

ABSTRACT

This disclosure relates to anti-cancer antibodies and methods of treatment, detection, and diagnosis using the same.

RELATED APPLICATIONS

This application is a 35 U.S.C. § 371 national stage filing ofInternational Application No. PCT/US2017/066661, filed Dec. 15, 2017,which in turuns claims the benefit of priority to U.S. ProvisionalPatent Application No. 62/435,509, filed on Dec. 16, 2016, U.S.Provisional Patent Application No. 62/488,445, filed on Apr. 21, 2017,and U.S. Provisional Patent Application No. 62/588,516, filed on Nov.20, 2017. The entire contents of each of the foregoing applications areincorporated herein by reference.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Nov. 29, 2017, isnamed 127913-00920_SL.txt and is 213,669 bytes in size.

BACKGROUND

CDCP1, also known as “CUB domain-containing protein 1,” “CD318,”“Transmembrane and associated with src kinases” (“TRASK”), and“SIMA135,” is an 836 amino acid, type I transmembrane protein thatconsists of a signal peptide of 29 amino acids, a larger extracellulardomain of 636 amino acids, heavily glycosylated with three regions thathave a low homology to C1r/C1s, urchin embryonic growth factor, and bonemor-phogenetic protein 1 (CUB) domains, a transmembrane and acytoplasmic domain of 21 and 150 amino acids, respectively. Thecytoplasmic domain of CDCP1 includes five conserved tyrosine residuesthat act as a substrate of Src Family Kinases (SFK) such as Src, Fyn andYes for subsequent phosphorylation.

CDCP1 is widely expressed in human epithelial tissues. CDCP1 functionsin the tyrosine phosphorylation-dependent regulation of cellular eventsthat are involved in tumor invasion and metastasis, but itsphosphorylation is only observed in mitotically detached or sheddingcells, consistent with its role in the negative regulation of celladhesion. The phosphorylation of CDCP1 is seen in many cancers,including some pre-invasive cancers as well as in invasive tumors and intumor metastases

Recent clinical and commercial success of anticancer antibodies hascreated great interest in antibody-based therapeutics. As antibodies cantarget specific antigens on the cancer cells, antibody-basedtherapeutics are likely to be more effective for treating cancers andhave lower toxic effects as compared to commonly-used chemotherapies.Thus, there is a need to develop anti-cancer antibodies for use invarious antibody-based therapeutics to treat cancers.

Antibody drug conjugates (ADC) represent a new class of therapeuticscomprising an antibody conjugated to a cytotoxic drug via a chemicallinker. The therapeutic concept of ADCs is to combine bindingcapabilities of an antibody with a drug, where the antibody is used todeliver the drug to a tumor cell by means of binding to a target surfaceantigen.

Accordingly, there remains a need in the art for anti-CDCP1 antibodiesand ADCs that can be used for therapeutic purposes in the treatment ofcancer.

SUMMARY

In certain aspects, the present invention provides for anti-CDCP1antibodies and antibody drug conjugates (ADCs). In certain embodimentsof the invention, the antibodies, or antigen binding portions thereof,bind to CDCP1 (e.g., the amino acid sequence provided in GenBankAccession No. NP_073753.3 and/or the amino acid sequence provided inNP_835488.1, the entire contects of each of which are incorporatedherein by reference), or the extracellular domain of CDCP1. In oneembodiment, the antibodies, or antigen binding portions thereof, of theinvention, bind to CDCP1 with a K_(d) of about 2,000 nM or less, about1,000 nM or less, about 500 nM or less, about 200 nM or less, about 100nM or less, about 75 nM or less, about 25 nM or less, about 21 nM orless, about 12 nM or less, about 11 nM or less, about 10 nM or less,about 9 nM or less, about 8 nM or less, about 7 nM or less, about 6 nMor less, about 5 nM or less, about 4 nM or less, about 3 nM or less,about 2 nM or less, about 1 nM or less, about 0.5 nM or less, about 0.3nM or less, about 0.1 nM or less, or about 0.01 nM or less, or about0.001 nM or less.

In yet other embodiments of the invention, anti-CDCP1 antibody drugconjugates (ADCs) of the invention (e.g., the CDCP1 antibodies of theinvention conjugated to a toxin) capable of being internalized. Inanother embodiment, the anti-CDCP1 antibody drug conjugates (ADCs) ofthe invention are capable of inducing cell death of cells endogenouslyexpressing CDCP1.

In one aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 157 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 160. Insome embodiments, the antibody, or antigen binding portion thereof,further comprises a heavy chain variable region comprising a CDR2 havingthe amino acid sequence of SEQ ID NO: 156 and a light chain variableregion comprising a CDR2 having the amino acid sequence of SEQ ID NO:159. In other embodiments, the antibody, or antigen binding portionthereof, comprises a heavy chain variable region comprising a CDR1having the amino acid sequence of SEQ ID NO: 155 and a light chainvariable region comprising a CDR1 having the amino acid sequence ofeither SEQ ID NO: 158.

In one aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 37 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 40. Insome embodiments, the antibody, or antigen binding portion thereof,further comprises a heavy chain variable region comprising a CDR2 havingthe amino acid sequence of SEQ ID NO: 36 and a light chain variableregion comprising a CDR2 having the amino acid sequence of SEQ ID NO:39. In other embodiments, the antibody, or antigen binding portionthereof, comprises a heavy chain variable region comprising a CDR1having the amino acid sequence of SEQ ID NO: 35 and a light chainvariable region comprising a CDR1 having the amino acid sequence ofeither SEQ ID NO: 38.

In one aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 87 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 90. Insome embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR2 having theamino acid sequence of SEQ ID NO: 86 and a light chain variable regioncomprising a CDR2 having the amino acid sequence of SEQ ID NO: 89. Inother embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR1 having theamino acid sequence of SEQ ID NO: 85 and a light chain variable regioncomprising a CDR1 having the amino acid sequence of either SEQ ID NO:88.

In yet another aspect of the invention, the present disclosure providesan isolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 107 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 110. Insome embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR2 having theamino acid sequence of SEQ ID NO: 106 and a light chain variable regioncomprising a CDR2 having the amino acid sequence of SEQ ID NO: 109. Inother embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR1 having theamino acid sequence of SEQ ID NO: 105 and a light chain variable regioncomprising a CDR1 having the amino acid sequence of either SEQ ID NO:108.

In another aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 127 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 130. Insome embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR2 having theamino acid sequence of SEQ ID NO: 126 and a light chain variable regioncomprising a CDR2 having the amino acid sequence of SEQ ID NO: 129. Inother embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR1 having theamino acid sequence of SEQ ID NO: 125 and a light chain variable regioncomprising a CDR1 having the amino acid sequence of either SEQ ID NO:128.

In one aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 47 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 50. Insome embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR2 having theamino acid sequence of SEQ ID NO: 46 and a light chain variable regioncomprising a CDR2 having the amino acid sequence of SEQ ID NO: 49. Inother embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR1 having theamino acid sequence of SEQ ID NO: 45 and a light chain variable regioncomprising a CDR1 having the amino acid sequence of either SEQ ID NO:48.

In another aspect of the invention, the present disclosure provides anisolated antibody, or antigen binding portion thereof, that binds tohuman CDCP1, wherein the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR3 having theamino acid sequence of SEQ ID NO: 77 and a light chain variable regioncomprising a CDR3 having the amino acid sequence of SEQ ID NO: 80. Insome embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR2 having theamino acid sequence of SEQ ID NO: 76 and a light chain variable regioncomprising a CDR2 having the amino acid sequence of SEQ ID NO: 79. Inother embodiments, the antibody, or antigen binding portion thereof,comprises a heavy chain variable region comprising a CDR1 having theamino acid sequence of SEQ ID NO: 75 and a light chain variable regioncomprising a CDR1 having the amino acid sequence of either SEQ ID NO:78.

In some aspects, the antibody, or antigen binding portion thereof, is anIgG isotype.

In some aspects, the antibody, or antigen binding portion thereof, has aK_(D) of 200 nM or less.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 157, a CDR2 domain comprising theamino acid sequence of SEQ ID NO: 156, and a CDR1 domain comprising theamino acid sequence of SEQ ID NO: 155, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 160, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:159, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:158.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 37, a CDR2 domain comprising the aminoacid sequence of SEQ ID NO: 36, and a CDR1 domain comprising the aminoacid sequence of SEQ ID NO: 35, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 40, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:39, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:38.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 87, a CDR2 domain comprising the aminoacid sequence of SEQ ID NO: 86, and a CDR1 domain comprising the aminoacid sequence of SEQ ID NO: 85, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 90, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:89, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:88.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 107, a CDR2 domain comprising theamino acid sequence of SEQ ID NO: 106, and a CDR1 domain comprising theamino acid sequence of SEQ ID NO: 105, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 110, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:109, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:108.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 127, a CDR2 domain comprising theamino acid sequence of SEQ ID NO: 126, and a CDR1 domain comprising theamino acid sequence of SEQ ID NO: 125, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 130, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:129, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:128.

In one aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 47, a CDR2 domain comprising the aminoacid sequence of SEQ ID NO: 46, and a CDR1 domain comprising the aminoacid sequence of SEQ ID NO: 45, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 50, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:49, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:48.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 77, a CDR2 domain comprising the aminoacid sequence of SEQ ID NO: 76, and a CDR1 domain comprising the aminoacid sequence of SEQ ID NO: 75, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 80, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:79, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:78.

In some aspects, the antibody, or antigen binding portion thereof, is anIgG isotype.

In some aspects, the antibody, or antigen binding portion thereof, has aK_(D) of 200 nM or less.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable domain comprising an amino acid sequence set forthin SEQ ID NO: 151 and a light chain variable domain comprising an aminoacid sequence set forth in SEQ ID NO: 152.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:151, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 151, and/or a light chain comprising an aminoacid sequence set forth in SEQ ID NO: 152, or a sequence having at least90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 152.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 31 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 32.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:31, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 31, and/or a light chain comprising an amino acidsequence set forth in SEQ ID NO: 32, or a sequence having at least 90%,95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 32.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 81 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 82.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:81, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 81, and/or a light chain comprising an amino acidsequence set forth in SEQ ID NO: 82, or a sequence having at least 90%,95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 82. In another aspectof the invention, the present disclosure provides an anti-CDCP1antibody, or antigen-binding portion thereof, comprising a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO: 101 anda light chain variable region comprising the amino acid sequence of SEQID NO: 102.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:101, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 101, and/or a light chain comprising an aminoacid sequence set forth in SEQ ID NO: 102, or a sequence having at least90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 102.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 121 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 122.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:121, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 121, and/or a light chain comprising an aminoacid sequence set forth in SEQ ID NO: 12, or a sequence having at least90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 122.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 41 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 42.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:41, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 41, and/or a light chain comprising an amino acidsequence set forth in SEQ ID NO: 42, or a sequence having at least 90%,95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 42.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 71 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 72.

In another aspect of the invention, the present disclosure provides ananti-CDCP1 antibody, or antigen-binding portion thereof, comprising aheavy chain comprising an amino acid sequence set forth in SEQ ID NO:71, or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99%identity to SEQ ID NO: 71, and/or a light chain comprising an amino acidsequence set forth in SEQ ID NO: 72, or a sequence having at least 90%,95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 72.

In one aspect of the invention, the present disclosure provides ananti-CUB domain-containing protein 1 (-CDCP1) antibody, orantigen-binding portion thereof, wherein the antibody, or antigenbinding portion thereof, comprises a heavy chain variable regioncomprising a CDR3 having an amino acid sequence selected from the groupconsisting of any one of the amino acid sequences in FIG. 15, 18 , or20, and a light chain variable region comprising a CDR3 having an aminoacid sequence selected from the group consisting of any one of the aminoacid sequences in FIG. 15, 18 , or 20.

In one embodiment, the heavy chain variable region further comprises aCDR2 having an amino acid sequence selected from the group consisting ofany one of the amino acid sequences in FIG. 15, 18 , or 20, and thelight chain variable region further comprises a CDR2 having an aminoacid sequence selected from the group consisting of any one of the aminoacid sequences in FIG. 15, 18 , or 20.

In another embodiment, the heavy chain variable region further comprisesa CDR1 having an amino acid sequence selected from the group consistingof any one of the amino acid sequences in FIG. 15, 18 , or 20, and thelight chain variable region further comprises a CDR1 having an aminoacid sequence selected from the group consisting of any one of the aminoacid sequences in FIG. 15, 18 , or 20.

In another aspect of the invention, the present disclosure provides anantibody, or antigen-binding portion thereof, that binds to the sameepitope as an antibody, or antigen-binding portion thereof, as describedherein.

In another aspect of the invention, the present disclosure provides anisolated nucleic acid encoding an antibody, or antigen binding portionthereof, as described herein.

In another aspect of the invention, the present disclosure provides apharmaceutical composition comprising the antibody, or antigen bindingportion thereof, as described herein, and a pharmaceutically acceptablecarrier.

In another aspect of the invention, the present disclosure provides anantibody, or antigen binding portion thereof, as described herein,conjugated to at least one drug.

In some aspects, the at least one drug is selected from the groupconsisting of an anti-apoptotic agent, a mitotic inhibitor, ananti-tumor antibiotic, an immunomodulating agent, a nucleic acid forgene therapy, an anti-angiogenic agent, an anti-metabolite, aboron-containing agent, a chemoprotective agent, a hormone agent, ananti-hormone agent, a corticosteroid, a photoactive therapeutic agent,an oligonucleotide, a radionuclide agent, a radiosensitizer, atopoisomerase inhibitor, and a tyrosine kinase inhibitor. In otherembodiments, the at least one drug is conjugated to the antibody, orantigen-binding portion thereof, via a linker. In another embodiment,the linker is a cleavable linker. In yet other embodiments, the linkeris a non-cleavable linker.

In one embodiment, the anti-CUB domain-containing protein 1 (-CDCP1)antibodies, or antigen-binding portions thereof, of the presentinvention are bispecific antibodies. In one embodiment, the anti-CUBdomain-containing protein 1 (-CDCP1) antibodies, or antigen-bindingportions thereof, of the present invention are multispecific antibodies.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 157, a CDR2 domain comprising the amino acid sequence of SEQID NO: 156, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 155, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 160, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 159, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 158.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 37, a CDR2 domain comprising the amino acid sequence of SEQID NO: 36, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 35, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 40, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 39, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 38.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 87, a CDR2 domain comprising the amino acid sequence of SEQID NO: 86, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 85, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 90, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 89, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 88.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 107, a CDR2 domain comprising the amino acid sequence of SEQID NO: 106, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 105, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 110, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 109, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 108.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 127, a CDR2 domain comprising the amino acid sequence of SEQID NO: 126, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 125, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 130, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 129, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 128.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 47, a CDR2 domain comprising the amino acid sequence of SEQID NO: 46, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 45, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 50, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 49, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 48.

In another aspect of the invention, the present disclosure provides anantibody drug conjugate (ADC) comprising an antibody, or antigen bindingportion thereof, conjugated to at least one drug, wherein the antibody,or antigen binding portion thereof, comprises a heavy chain variableregion comprising a CDR3 domain comprising the amino acid sequence ofSEQ ID NO: 77, a CDR2 domain comprising the amino acid sequence of SEQID NO: 76, and a CDR1 domain comprising the amino acid sequence of SEQID NO: 75, and a light chain variable region comprising a CDR3 domaincomprising the amino acid sequence of SEQ ID NO: 80, a CDR2 domaincomprising the amino acid sequence of SEQ ID NO: 79, and a CDR1 domaincomprising the amino acid sequence of SEQ ID NO: 78.

In another aspect of the invention, the present disclosure provides anantibody, or antigen binding portion thereof, as described herein,conjugated to at least one drug.

In some embodiments, the at least one drug is selected from the groupconsisting of an anti-apoptotic agent, a mitotic inhibitor, ananti-tumor antibiotic, an immunomodulating agent, a nucleic acid forgene therapy, an anti-angiogenic agent, an anti-metabolite, aboron-containing agent, a chemoprotective agent, a hormone agent, ananti-hormone agent, a corticosteroid, a photoactive therapeutic agent,an oligonucleotide, a radionuclide agent, a radiosensitizer, atopoisomerase inhibitor, and a tyrosine kinase inhibitor. In otherembodiments, the at least one drug is conjugated to the antibody, orantigen-binding portion thereof, via a linker. In yet other embodiments,the linker is a cleavable linker. In another embodiment, the linker is anon-cleavable linker.

In some embodiments, the at least one drug is conjugated via a linker.In other embodiments, the linker is a cleavable linker. In yet otherembodiments, the linker is a non-cleavable linker.

In some embodiments, the antibody, or antigen binding portion thereof,is an IgG1 isotype.

In another aspect of the invention, the present disclosure provides apharmaceutical composition comprising an ADC mixture comprising aplurality of the ADC as described herein, and a pharmaceuticallyacceptable carrier.

In some embodiments, the ADC mixture has an average drug to antibodyratio (DAR) of 0 to 8.

In another aspect of the invention, the present disclosure provides amethod for treating cancer, comprising administering a therapeuticallyeffective amount of an antibody or antigen binding portion thereof, asdescribed herein, or a bispecific antibody as described herein, or anADC as described herein, to a subject in need thereof.

In some embodiments, the cancer is selected from the group consisting ofbreast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer. Inone embodiment, the cancer is breast cancer, e.g., triple negativebreast cancer. In one embodiment, the cancer is colon cancer. In oneembodiment, the cancer is lung cancer, e.g., non-small cell lung cancer(NSCLC).

In another aspect, the present invention provides a method forinhibiting or decreasing solid tumor growth in a subject having a solidtumor. The method includes administering a therapeutically effectiveamount of an antibody or antigen binding portion thereof, as describedherein, or a bispecific antibody as described herein, or an ADC asdescribed herein to the subject having the solid tumor, such that thesolid tumor growth is inhibited or decreased.

In some embodiments, the cancer is selected from the group consisting ofbreast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer. Inone embodiment, the cancer is breast cancer, e.g., triple negativebreast cancer. In one embodiment, the cancer is colon cancer. In oneembodiment, the cancer is lung cancer, e.g., non-small cell lung cancer(NSCLC).

In one embodiment, the antibody or antigen binding portion thereof, or abispecific antibody as described herein, or the ADC is administered incombination with an additional agent or an additional therapy. In oneembodiment, the additional agent is an immune checkpoint inhibitor,e.g., an antibody, such as an antibody selected from the groupconsisting of an anti-PD1 antibody, an anti-PD-L1 antibody and ananti-CTLA-4 antibody. In one embodiment, the additional therapy isradiation. In one embodiment, the additional agent is a chemotherapeuticagent.

In one embodiment, the cancer or tumor is characterized as having CDCP1expression or overexpression.

In another aspect of the invention, the present disclosure provides amethod for treating cancer. The method includes administering atherapeutically effective amount of of an antibody or antigen bindingportion thereof, as described herein, or a bispecific antibody asdescribed herein, or an ADC as described herein, and an anti-PD-L1antibody to a subject in need thereof.

In some embodiments, the cancer is selected from the group consisting ofbreast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer. Inone embodiment, the cancer is breast cancer, e.g., triple negativebreast cancer. In one embodiment, the cancer is colon cancer. In oneembodiment, the cancer is lung cancer, e.g., non-small cell lung cancer(NSCLC).

In another aspect, the present invention provides a method forinhibiting or decreasing solid tumor growth in a subject having a solidtumor. The method includes administering a therapeutically effectiveamount of an antibody or antigen binding portion thereof, as describedherein, or a bispecific antibody as described herein, or an ADC asdescribed herein, and an anti-PD-L1 antibody to the subject having thesolid tumor, such that the solid tumor growth is inhibited or decreased.

In some embodiments, the cancer is selected from the group consisting ofbreast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer. Inone embodiment, the cancer is breast cancer, e.g., triple negativebreast cancer. In one embodiment, the cancer is colon cancer. In oneembodiment, the cancer is lung cancer, e.g., non-small cell lung cancer(NSCLC).

In one embodiment, the antibody or antigen binding portion thereof or abispecific antibody as described herein, or the ADC is administered incombination with an additional agent or an additional therapy. In oneembodiment, the additional agent is an immune checkpoint inhibitor,e.g., an antibody, such as an antibody that is selected from the groupconsisting of an anti-PD1 antibody and an anti-CTLA-4 antibody. In oneembodiment, the additional therapy is radiation. In one embodiment, theadditional agent is a chemotherapeutic agent.

In one embodiment, the cancer or tumor is characterized as having CDCP1expression or overexpression.

In some embodiments, the present disclosure provides a method forinhibiting or decreasing solid tumor growth in a subject having a solidtumor, said method comprising administering an effective amount of theantibody or antigen binding portion thereof, as described herein, or theADC, as described herein, to the subject having the solid tumor, suchthat the solid tumor growth is inhibited or decreased.

In some embodiments, the antibody or antigen binding portion thereof orthe ADC is administered in combination with an additional agent or anadditional therapy. In other embodiments, the additional agent is animmune checkpoint inhibitor. In yet another embodiment, the immunecheckpoint inhibitor is an antibody. In another embodiment, the antibodyis selected from the group consisting of an anti-PD1 antibody, ananti-PD-L1 antibody or an anti-CTLA-4 antibody. In other embodiments,the additional therapy is radiation. In yet another embodiment, theadditional agent is a chemotherapeutic agent. In some embodiments, thecancer or tumor is characterized as having CDCP1 expression oroverexpression.

In another aspect, the disclosure relates to a polynucleotide linked toa heterologous nucleic acid, wherein the polynucleotide is selected fromthe group consisting of (a) a polynucleotide encoding an immunoglobulinheavy chain or a fragment thereof comprising a heavy chain variableregion (VH) comprising complementarity determining regions (CDRs) 1, 2,and 3 with the amino acid sequences set forth in FIG. 20 , and whereinthe VH when paired with the corresponding light chain variable region(VL) as shown in FIG. 15 binds to CDCP1 protein; (b) a polynucleotideencoding an immunoglobulin light chain or a fragment thereof comprisinga VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forthin FIG. 20 , and wherein the VL when paired with the corresponding VH asshown in FIG. 15 binds to CDCP1; (c) a polynucleotide encoding (i) animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inFIG. 20 ; and (ii) an immunoglobulin light chain or a fragment thereofcomprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in FIG. 20 ; and (d) a polynucleotide encoding animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising the amino acid sequence set forth in FIG. 18 , wherein the VHwhen paired with the corresponding VL as shown in FIG. 15 binds toCDCP1.

In one aspect, the disclosure features an expression vector comprising aheterologous promoter operably linked to a polynucleotide encoding apolypeptide disclosed herein.

In another aspect, the disclosure provides an expression vectorcomprising: a first polynucleotide encoding a first polypeptidecomprising an immunoglobulin heavy chain or a fragment thereofcomprising a heavy chain variable region (VH) comprising VHcomplementarity determining regions (CDRs) 1, 2, and 3 with the aminoacid sequences set forth in FIG. 20 ; and a second polynucleotideencoding a second polypeptide comprising an immunoglobulin light chainor a fragment thereof comprising a light chain variable region (VL)comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forthin FIG. 20 , wherein the immunoglobulin heavy chain or fragment thereofwhen paired with the immunoglobulin light chain or fragment thereofforms an anti-CDCP1 antibody or CDCP1-binding fragment thereof, andwherein the expression vector is, for example, a plasmid, phage, orvirus.

In some embodiments, the complementarity determining region (CDR)sequences in the heavy chain variable region and the light chainvariable region comprise or consist of the CDR sequences as set forth inFIG. 20 . In some embodiments, the CDR sequences in the heavy chainvariable region and the light chain variable region are at least 80%,90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to the CDR sequences asset forth in FIG. 20 . In some embodiments, the CDR sequences in theheavy chain variable region and the light chain variable region differfrom the CDR sequences in FIG. 20 by one, two, three, four, or fiveamino acids. In some embodiments, the CDRs in FIG. 20 are substituted byone, two, three, four, or five conservative amino acids.

In another aspect, the present disclosure features compositions thatinclude at least one antibody disclosed herein conjugated to atherapeutic agent, e.g., a cytotoxic drug or radioisotope, and/or to areporter group. In some embodiments, an antibody that is conjugated to atherapeutic agent is administered to a subject to induce cell death of acancer cell, e.g., a cancer cell that expresses or overexpresses CDCP1protein, or has CDCP1 protein on the cell surface.

In some embodiments, the antibodies are conjugated to a detectablemarker. The conjugates are administered to a subject to detect a cancercell that expresses CDCP1 protein and/or has a detectable and/orelevated level of a CDCP1 protein.

In some embodiments, an antibody is linked (e.g., covalently bonded,hydrogen bonded, or ionicly bonded) to a surface (e.g., a microfluidicdevice, a chromatography resin, an array, polymer, or a bead).

In a further aspect, the disclosure features compositions that includeat least one of the antibodies disclosed herein and a pharmaceuticallyacceptable excipient. In some embodiments, the disclosure features dry(e.g., lyophilized) compositions that include one or more antibodiesdisclosed herein and, optionally, one or more pharmaceuticallyacceptable excipients.

In another aspect, the disclosure features polynucleotides, e.g., DNA,that encode a polypeptide chain, e.g., an antibody heavy or light chain,of any of the antibodies disclosed herein. For example, thepolynucleotide may include a sequence disclosed herein. In someembodiments, the polynucleotides, e.g., DNA, do not include introns. Thedisclosure also features vectors, e.g., recombinant vectors andexpression vectors that include the above polynucleotides, and a cell,e.g., an isolated cell, e.g., recombinant cells or hybridomas, thatinclude the above polynucleotides and/or vectors. In some embodiments,the vector is stably integrated into a chromosome of the cell, e.g., amammalian cell, bacterial cell, or yeast cell. In some embodiments, thedisclosure features methods of producing antibodies that includeculturing the cells, e.g., isolated cells, under conditions where theantibodies are expressed, and methods of collecting the antibodies.

The disclosure features the antibodies, nucleic acids, compositions, andcells disclosed herein and the use thereof for treatment, prophylaxis,imaging, and/or diagnosis of a cancer. In some embodiments, thecomposition is formulated for intravenous administration. In someembodiments, the cancer expresses (e.g., overexpresses) CDCP1 or hasCDCP1 on the cell surface. In some embodiments, the cancer ischaracterized by the presence of a CDCP1 and/or an elevated level of aCDCP1 protein (e.g., as compared to a reference level, e.g., a level ofa CDCP1 protein in a CDCP1 protein produced by a healthy subject)produced by the cancer cells. In a further aspect, the disclosurefeatures methods for treatment of a cancer (e.g., a cancer characterizedby overexpression of CDCP1 in cancer cells, or a cancer characterized byhaving CDCP1 on the surface of the cancer cells) (e.g., breast cancer,triple-negative breast cancer, carcinoid cancer, cervical cancer,endometrial cancer, glioma, head and neck cancer, liver cancer, lungcancer, small cell lung cancer, lymphoma, melanoma, ovarian cancer,pancreatic cancer, prostate cancer, renal cancer, gastric cancer,testicular cancer, thyroid cancer, bladder cancer, urethral cancer,colorectal cancer, or hematologic malignancy), that includeadministering an antibody, nucleic acid, composition, or cell disclosedherein to a subject with a cancer in a therapeutically effective amount.

In another aspect, the disclosure features methods that includeadministering an antibody or composition, e.g., a cell composition,antibody-drug conjugate, or antibody-radioisotope conjugate disclosedherein to a subject in need thereof, e.g., a subject having, oridentified or diagnosed as having a cancer characterized byoverexpression of CDCP1 in cancer cells, or a cancer characterized byhaving CDCP1 on the surface of the cancer cells, e.g., breast cancer,triple-negative breast cancer, carcinoid cancer, cervical cancer,endometrial cancer, glioma, head and neck cancer, liver cancer, lungcancer, small cell lung cancer, non-small cell lung cancer, lymphoma,melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renalcancer, gastric cancer, testicular cancer, thyroid cancer, bladdercancer, urethral cancer, colorectal cancer, colon cancer, or hematologicmalignancy cells.

In some embodiments, the subject is identified as being a subject whoexpresses CDCP1, e.g., using any of the methods described herein, or hasan elevated level of a CDCP1 protein, e.g., as compared to a referencelevel, e.g., a level of a CDCP1 protein in a CDCP1 protein produced by ahealthy subject, a level of a CDCP1 protein in CDCP1 protein produced bya non-cancerous, e.g., primary cell, or a threshold level of a CDCP1protein, in which a determined level of a CDCP1 protein that is abovethis value indicates that the subject should be administered an antibodydescribed herein.

In yet another aspect, the disclosure features methods for cancerprophylaxis (or reducing a subject's risk of developing a cancercharacterized by expression, e.g., overexpression, of CDCP1 protein incancer cells or a cancer characterized by having CDCP1 on the surface ofthe cancer cells, e.g., breast cancer (e.g., triple-negative breastcancer), carcinoid cancer, cervical cancer, endometrial cancer, glioma,head and neck cancer, liver cancer, lung cancer, small cell lung cancer,lymphoma, melanoma, ovarian cancer, pancreatic cancer, prostate cancer,renal cancer, gastric cancer, testicular cancer, thyroid cancer, bladdercancer, urethral cancer, colorectal cancer, or hematologic malignancycells (e.g., as compared to a subject at risk for developing cancer butreceiving no treatment or receiving a different treatment). The methodsinclude administering an antibody, nucleic acid, composition, or celldisclosed herein to a subject in need thereof in a prophylacticallyeffective amount. In some embodiments, the cancer expresses CDCP1protein. In some embodiments, the cancer cells have CDCP1 on the cellsurface. In some embodiments of any of these methods, the subject isidentified as having an elevated risk of developing cancer.

In another aspect, the disclosure features methods of detecting a CDCP1protein (e.g., a CDCP1 protein) in a sample (e.g., a biopsy sample). Themethods include contacting a sample with an antibody disclosed hereinand detecting binding of the agent to the sample, thereby detectingCDCP1 protein in the sample. Some embodiments further include recordingthe detection or non-detection of CDCP1 protein in the clinical recordsof a subject from whom the sample was obtained. In some embodiments, theclinical record is stored on a tangible computer readable medium, e.g.,a disc, magnetic tape, or computer memory.

In some embodiments of any of the methods described herein, theantibodies described herein are contacted with a sample and/or a cell,and the antibody to CDCP1 can be used in an immunoassay (e.g., anenzyme-linked immunosorbent assay), fluorescence-assisted cell sorting,microfluidics, and chromatography.

In one aspect, the present invention provides an antibody orantigen-binding fragment thereof that binds to CUB domain-containingprotein 1 (CDCP1). The antibody or antigen-binding fragment thereofcomprises a heavy chain variable region (VH) comprising complementaritydetermining regions (CDRs) 1, 2, 3, wherein the CDR1 region comprises anamino acid sequence that is at least 80% identical to a selected VH CDR1amino acid sequence, the CDR2 region comprises an amino acid sequencethat is at least 80% identical to a selected VH CDR2 amino acidsequence, and the CDR3 region comprises an amino acid sequence that isat least 80% identical to a selected VH CDR3 amino acid sequence, and alight chain variable region (VL) comprising CDRs 1, 2, 3, wherein theCDR1 region comprises an amino acid sequence that is at least 80%identical to a selected VL CDR1 amino acid sequence, the CDR2 regioncomprises an amino acid sequence that is at least 80% identical to aselected VL CDR2 amino acid sequence, and the CDR3 region comprises anamino acid sequence that is at least 80% identical to a selected VL CDR3amino acid sequence, wherein the selected VH CDRs 1, 2, 3 amino acidsequences and the selected VL CDRs, 1, 2, 3 amino acid sequences are oneof the following: (1) the selected VH CDRs 1, 2, 3 amino acid sequencesare set forth in SEQ ID NOs: 5, 6, 7, respectively, and the selected VLCDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 8, 9, 10,respectively; (2) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 15, 16, 17, respectively, and the selected VLCDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 18, 19,20, respectively; (3) the selected VH CDRs 1, 2, 3 amino acid sequencesare set forth in SEQ ID NOs: 25, 26, 27, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 28,29, 30, respectively; (4) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 35, 36, 37, respectively, and theselected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ IDNOs: 38, 39, 40, respectively; (5) the selected VH CDRs 1, 2, 3 aminoacid sequences are set forth in SEQ ID NOs: 45, 46, 47, respectively,and the selected VL CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 48, 49, 50, respectively; (6) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 55, 56, 57,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 58, 59, 60, respectively; (7) the selected VHCDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 65, 66,67, respectively, and the selected VL CDRs 1, 2, 3 amino acid sequencesare set forth in SEQ ID NOs: 68, 69, 70, respectively; (8) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 75,76, 77, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 78, 79, 80, respectively; (9) theselected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ IDNOs: 85, 86, 87, respectively, and the selected VL CDRs 1, 2, 3 aminoacid sequences are set forth in SEQ ID NOs: 88, 89, 90, respectively;(10) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 95, 96, 97, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 98, 99, 100,respectively; (11) the selected VI-1 CDRs 1, 2, 3 amino acid sequencesare set forth in SEQ ID NOs: 105, 106, 107, respectively, and theselected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ IDNOs: 108, 109, 110, respectively; (12) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 115, 116, 117,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 118, 119, 120, respectively; (13) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 125,126, 127, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 128, 129, 130, respectively; (14)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 135, 136, 137, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 138, 139, 140,respectively; (15) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 145, 146, 147, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 148,149, 150, respectively; (16) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 155, 156, 157, respectively, andthe selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 158, 159, 160, respectively; (17) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 165, 166, 167,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 168, 169, 170, respectively; (18) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 175,176, 177, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 178, 179, 180, respectively; (19)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 185, 186, 187, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 188, 189, 190,respectively; (20) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 195, 196, 197, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 198,199, 200, respectively; (21) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 205, 206, 207, respectively, andthe selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 208, 209, 210, respectively; (22) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 215, 216, 217,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 218, 219, 220, respectively; (23) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 225,226, 227, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 228, 229, 230, respectively; (24)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 235, 236, 237, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 238, 239, 240,respectively; (25) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 245, 246, 247, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 248,249, 250, respectively; (26) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 255, 256, 257, respectively, andthe selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 258, 259, 260, respectively; (27) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 265, 266, 267,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 268, 269, 270, respectively; (28) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 275,276, 277, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 278, 279, 280, respectively; (29)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 285, 286, 287, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 288, 289, 290,respectively; (30) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 295, 296, 297, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 298,299, 300, respectively; (31) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 305, 306, 307, respectively, andthe selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 308, 309, 310, respectively; (32) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 315, 316, 317,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ 113 NOs: 318, 319, 320, respectively; (33) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 325,326, 327, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 328, 329, 330, respectively; (34)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 335, 336, 337, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 338, 339, 340,respectively; (35) the selected VH CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 345, 346, 347, respectively, and the selectedVL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 348,349, 350, respectively; (36) the selected VH CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 355, 356, 357, respectively, andthe selected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 358, 359, 360, respectively; (37) the selected VH CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 365, 366, 367,respectively, and the selected VL CDRs 1, 2, 3 amino acid sequences areset forth in SEQ ID NOs: 368, 369, 370, respectively; (38) the selectedVH CDRs 1, 2, 3 amino acid sequences are set forth in SEQ ID NOs: 375,376, 377, respectively, and the selected VL CDRs 1, 2, 3 amino acidsequences are set forth in SEQ ID NOs: 378, 379, 380, respectively; (39)the selected VH CDRs 1, 2, 3 amino acid sequences are set forth in SEQID NOs: 385, 386, 387, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 388, 389, 390,respectively; and (40) the selected VH CDRs 1, 2, 3 amino acid sequencesare set forth in SEQ ID NOs: 395, 396, 397, respectively, and theselected VL CDRs 1, 2, 3 amino acid sequences are set forth in SEQ IDNOs: 398, 399, 400, respectively, wherein the antibody orantigen-binding fragment thereof specifically binds to CUBdomain-containing protein 1 (CDCP1).

In one embodiment, the antibody or antigen-binding fragment thereofspecifically binds to human CDCP1 and/or Cynomolgus CDCP1. In oneembodiment, the antibody or antigen-binding fragment thereof has adissociation constant (K_(a)) for human CDCP1 that is less than 10 nM,and/or a K_(a) for Cynomolgus CDCP1 that is less than 10 nM. In oneembodiment, the antibody or antigen-binding fragment thereof is ahumanized antibody or antigen-binding fragment thereof.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 5, 6, and 7 respectively, and the VL comprises CDRs 1, 2,3 with the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10,respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 15, 16, and 17 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 18, 19, and20, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 25, 26, and 27 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 28, 29, and30, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 35, 36, and 37 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 38, 39, and40, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 45, 46, and 47 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 48, 49, and50, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 55, 56, and 57 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 58, 59, and60, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 65, 66, and 67 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 68, 69, and70, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 75, 76, and 77 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 78, 79, and80, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 85, 86, and 87 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 88, 89, and90, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 95, 96, and 97 respectively, and the VL comprises CDRs 1,2, 3 with the amino acid sequences set forth in SEQ ID NOs: 98, 99, and100, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 105, 106, and 107 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 108, 109,and 110, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 115, 116, and 117 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 118, 119,and 120, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 125, 126, and 127 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 128, 129,and 130, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 135, 136, and 137 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 138, 139,and 140, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 145, 146, and 147 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 148, 149,and 150, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 155, 156, and 157 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 158, 159,and 160, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 165, 166, and 167 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 168, 169,and 170, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 175, 176, and 177 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 178, 179,and 180, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 185, 186, and 187 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 188, 189,and 190, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 195, 196, and 197 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 198, 199,and 200, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 205, 206, and 207 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 208, 209,and 210, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 215, 216, and 217 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 218, 219,and 220, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 225, 226, and 227 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 228, 229,and 230, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 235, 236, and 237 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 238, 239,and 240, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 245, 246, and 247 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 248, 249,and 250, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 255, 256, and 257 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 258, 259,and 260, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 265, 266, and 267 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 268, 269,and 270, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 275, 276, and 277 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 278, 279,and 280, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 285, 286, and 287 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 288, 289,and 290, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 295, 296, and 297 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 298, 299,and 300, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 305, 306, and 307 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 308, 309,and 310, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 315, 316, and 317 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 318, 319,and 320, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 325, 326, and 327 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 328, 329,and 330, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 335, 336, and 337 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 338, 339,and 340, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 345, 346, and 347 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 348, 349,and 350, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 355, 356, and 357 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 358, 359,and 360, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 365, 366, and 367 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 368, 369,and 370, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 375, 376, and 377 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 378, 379,and 380, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 385, 386, and 387 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 388, 389,and 390, respectively.

In one embodiment, the VH of the antibody or antigen-binding fragmentthereof comprises CDRs 1, 2, 3 with the amino acid sequences set forthin SEQ ID NOs: 395, 396, and 397 respectively, and the VL comprises CDRs1, 2, 3 with the amino acid sequences set forth in SEQ ID NOs: 398, 399,and 400, respectively.

In another aspect, the present invention provides a cDNA comprising apolynucleotide encoding a polypeptide comprises (1) an immunoglobulinheavy chain or a fragment thereof comprising a heavy chain variableregion (VH) comprising complementarity determining regions (CDRs) 1, 2,and 3 with the amino acid sequences set forth in SEQ ID NOs: 5, 6, and7, respectively, and wherein the VH when paired with a light chainvariable region (VL) comprising the amino acid sequence set forth in SEQID NO: 2 binds to CDCP1; (2) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 8, 9, and 10, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO:1 binds to CDCP1; (3) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 15, 16, and 17,respectively, and wherein the VH when paired with a VL comprising theamino acid sequence set forth in SEQ ID NO: 12 binds to CDCP1; (4) animmunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 18, 19, and 20, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 11binds to CDCP1; (5) an immunoglobulin heavy chain or a fragment thereofcomprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 25, 26, and 27, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 22 binds to CDCP1; (6) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 28, 29, and 30,respectively, and wherein the VL when paired with a VH comprising theamino acid sequence set forth in SEQ ID NO: 21 binds to CDCP1; (7) animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 35, 36, and 37, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 32binds to CDCP1; (8) an immunoglobulin light chain or a fragment thereofcomprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 38, 39, and 40, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 31 binds to CDCP1; (9) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 45, 46, and 47,respectively, and wherein the VH when paired with a VL comprising theamino acid sequence set forth in SEQ ID NO: 42 binds to CDCP1; (10) animmunoglobulin light chain ora fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 48, 49, and 50, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 41binds to CDCP1; (11) an immunoglobulin heavy chain or a fragment thereofcomprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 55, 56, and 57, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 52 binds to CDCP1; (12) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 58, 59, and 60,respectively, and wherein the VL when paired with a VH comprising theamino acid sequence set forth in SEQ ID NO: 51 binds to CDCP1; (13) animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 65, 66, and 67, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 62binds to CDCP1; (14) an immunoglobulin light chain or a fragment thereofcomprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 68, 69, and 70, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 61 binds to CDCP1; (15) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 75, 76, and 77,respectively, and wherein the VH when paired with a VL comprising theamino acid sequence set forth in SEQ ID NO: 72 binds to CDCP1; (16) animmunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 78, 79, and 80, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 71binds to CDCP1; (17) an immunoglobulin heavy chain or a fragment thereofcomprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 85, 86, and 87, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 82 binds to CDCP1; (18) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 88, 89, and 90,respectively, and wherein the VL when paired with a VH comprising theamino acid sequence set forth in SEQ ID NO: 81 binds to CDCP1; (19) animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 95, 96, and 97, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 92binds to CDCP1; (20) an immunoglobulin light chain or a fragment thereofcomprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 98, 99, and 100, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 91 binds to CDCP1; (21) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 105, 106, and107, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 102 binds to CDCP1; (22)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 108, 109, and 110, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 101 binds to CDCP1; (23) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 115, 116, and 117, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 112 binds to CDCP1; (24) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 118, 119, and120, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 111 binds to CDCP1; (25)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 125, 126, and 127, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 122 binds to CDCP1; (26) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 128, 129, and 130, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 121 binds to CDCP1; (27) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 135, 136, and137, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 132 binds to CDCP1; (28)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 138, 139, and 140, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 131 binds to CDCP1; (29) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 145, 146, and 147, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 142 binds to CDCP1; (30) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 148, 149, and150, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 141 binds to CDCP1; (31)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 155, 156, and 157, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 152 binds to CDCP1; (32) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 158, 159, and 160, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 151 binds to CDCP1; (33) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 165, 166, and167, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 162 binds to CDCP1; (34)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 168, 169, and 170, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 161 binds to CDCP1; (35) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 175, 176, and 177, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 172 binds to CDCP1; (36) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 178, 179, and180, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 171 binds to CDCP1; (37)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 185, 186, and 187, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 182 binds to CDCP1; (38) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 188, 189, and 190, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 181 binds to CDCP1; (39) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 195, 196, and197, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 192 binds to CDCP1; (40)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 198, 199, and 200, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 191 binds to CDCP1; (41) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 205, 206, and 207, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 202 binds to CDCP1; (42) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 208, 209, and210, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 201 binds to CDCP1; (43)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 215, 216, and 217, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 212 binds to CDCP1; (44) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 218, 219, and 220, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 211 binds to CDCP1; (45) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 225, 226, and227, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 222 binds to CDCP1; (46)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 228, 229, and 230, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 221 binds to CDCP1; (47) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 235, 236, and 237, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 232 binds to CDCP1; (48) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 238, 239, and240, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 231 binds to CDCP1; (49)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 245, 246, and 247, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 242 binds to CDCP1; (50) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 248, 249, and 250, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 241 binds to CDCP1; (51) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 255, 256, and257, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 252 binds to CDCP1; (52)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 258, 259, and 260, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 251 binds to CDCP1; (53) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 265, 266, and 267, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 262 binds to CDCP1; (54) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 268, 269, and270, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 261 binds to CDCP1; (55)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 275, 276, and 277, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 272 binds to CDCP1; (56) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 278, 279, and 280, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 271 binds to CDCP1; (57) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 285, 286, and287, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 282 binds to CDCP1; (58)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 288, 289, and 290, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 281 binds to CDCP1; (59) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 295, 296, and 297, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 292 binds to CDCP1; (60) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 298, 299, and300, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 291 binds to CDCP1; (61)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 305, 306, and 307, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 302 binds to CDCP1; (62) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 308, 309, and 310, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 301 binds to CDCP1; (63) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 315, 316, and317, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 312 binds to CDCP1; (64)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 318, 319, and 320, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 311 binds to CDCP1; (65) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 325, 326, and 327, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 322 binds to CDCP1; (66) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 328, 329, and330, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 321 binds to CDCP1; (67)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 335, 336, and 337, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 332 binds to CDCP1; (68) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 338, 339, and 340, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 331 binds to CDCP1; (69) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 345, 346, and347, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 342 binds to CDCP1; (70)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 348, 349, and 350, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 341 binds to CDCP1; (71) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 355, 356, and 357, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 352 binds to CDCP1; (72) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 358, 359, and360, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 351 binds to CDCP1; (73)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 365, 366, and 367, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 362 binds to CDCP1; (74) an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 368, 369, and 370, respectively, andwherein the VL when paired with a VH comprising the amino acid sequenceset forth in SEQ ID NO: 361 binds to CDCP1; (75) an immunoglobulin heavychain or a fragment thereof comprising a VH comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 375, 376, and377, respectively, and wherein the VH when paired with a VL comprisingthe amino acid sequence set forth in SEQ ID NO: 372 binds to CDCP1; (76)an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 378, 379, and 380, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 371 binds to CDCP1; (77) an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 385, 386, and 387, respectively, andwherein the VH when paired with a VL comprising the amino acid sequenceset forth in SEQ ID NO: 382 binds to CDCP1; (78) an immunoglobulin lightchain or a fragment thereof comprising a VL comprising CDRs 1, 2, and 3with the amino acid sequences set forth in SEQ ID NOs: 388, 389, and390, respectively, and wherein the VL when paired with a VH comprisingthe amino acid sequence set forth in SEQ ID NO: 381 binds to CDCP1; (79)an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 395, 396, and 397, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 392 binds to CDCP1; or (80) an immunoglobulin light chain or afragment thereof comprising a VL comprising CDRs 1, 2, and 3 with theamino acid sequences set forth in SEQ ID NOs: 398, 399, and 400,respectively, and wherein the VL when paired with a VH comprising theamino acid sequence set forth in SEQ ID NO: 391 binds to CDCP1.

In one embodiment, the VH when paired with a VL specifically binds tohuman CDCP1 and/or Cynomolgus CDCP1, and the VL when paired with a VHspecifically binds to human CDCP1 and/or Cynomolgus CDCP1.

In one embodiment, the immunoglobulin heavy chain or the fragmentthereof is a humanized immunoglobulin heavy chain or a fragment thereof,and the immunoglobulin light chain or the fragment thereof is ahumanized immunoglobulin light chain or a fragment thereof.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 5, 6, and 7, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 8, 9, and 10, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 15, 16, and 17, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 18, 19, and 20, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 25, 26, and 27, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 28, 29, and 30, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 35, 36, and 37, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 38, 39, and 40, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 45, 46, and 47, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 48, 49, and 50, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 55, 56, and 57, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 58, 59, and 60, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 65, 66, and 67, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 68, 69, and 70, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 75, 76, and 77, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 78, 79, and 80, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 85, 86, and 87, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 88, 89, and 90, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 95, 96, and 97, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 98, 99, and 100, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 105, 106, and 107, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 108, 109, and 110, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 115, 116, and 117, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 118, 119, and 120, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 125, 126, and 127, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 128, 129, and 130, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 135, 136, and 137, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 138, 139, and 140, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 145, 146, and 147, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 148, 149, and 150, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 155, 156, and 157, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 158, 159, and 160, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 165, 166, and 167, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 168, 169, and 170, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 175, 176, and 177, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 178, 179, and 180, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 185, 186, and 187, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 188, 189, and 190, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 195, 196, and 197, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 198, 199, and 200, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 205, 206, and 207, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 208, 209, and 210, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 215, 216, and 217, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 218, 219, and 220, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 225, 226, and 227, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 228, 229, and 230, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 235, 236, and 237, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 238, 239, and 240, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 245, 246, and 247, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 248, 249, and 250, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 255, 256, and 257, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 258, 259, and 260, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 265, 266, and 267, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 268, 269, and 270, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 275, 276, and 277, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 278, 279, and 280, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 285, 286, and 287, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 288, 289, and 290, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 295, 296, and 297, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 298, 299, and 300, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 305, 306, and 307, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 308, 309, and 310, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 315, 316, and 317, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 318, 319, and 320, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 325, 326, and 327, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 328, 329, and 330, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 335, 336, and 337, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 338, 339, and 340, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 345, 346, and 347, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 348, 349, and 350, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 355, 356, and 357, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 358, 359, and 360, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 365, 366, and 367, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 368, 369, and 370, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 375, 376, and 377, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 378, 379, and 380, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 385, 386, and 387, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 388, 389, and 390, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin heavy chain or a fragmentthereof comprising a VH comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 395, 396, and 397, respectively.

In one embodiment, the cDNA comprises a polynucleotide encoding apolypeptide comprising an immunoglobulin light chain or a fragmentthereof comprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in SEQ ID NOs: 398, 399, and 400, respectively.

In one aspect, the present invention provides an antibody orantigen-binding fragment thereof binds to CDCP1 comprising a heavy chainvariable region (VH) comprising an amino acid sequence that is at least90% identical to a selected VH sequence, and a light chain variableregion (VL) comprising an amino acid sequence that is at least 90%identical to a selected VL sequence, wherein the selected VH sequenceand the selected VL sequence are one of the following: (1) the selectedVH sequence is SEQ ID NO: 1, and the selected VL sequence is SEQ ID NO:2; (2) the selected VH sequence is SEQ ID NO: 11, and the selected VLsequence is SEQ ID NO: 12; (3) the selected VH sequence is SEQ ID NO:21, and the selected VL sequence is SEQ ID NO: 22; (4) the selected VHsequence is SEQ ID NO: 31, and the selected VL sequence is SEQ ID NO:32; (5) the selected VH sequence is SEQ ID NO: 41, and the selected VLsequence is SEQ ID NO: 42; (6) the selected VH sequence is SEQ ID NO:51, and the selected VL sequence is SEQ ID NO: 52; (7) the selected VHsequence is SEQ ID NO: 61, and the selected VL sequence is SEQ ID NO:62; (8) the selected VH sequence is SEQ ID NO: 71, and the selected VLsequence is SEQ ID NO: 72; (9) the selected VH sequence is SEQ ID NO:81, and the selected VL sequence is SEQ ID NO: 82; (10) the selected VHsequence is SEQ ID NO: 91, and the selected VL sequence is SEQ ID NO:92; (11) the selected VH sequence is SEQ ID NO: 101, and the selected VLsequence is SEQ ID NO: 102; (12) the selected VH sequence is SEQ ID NO:111, and the selected VL sequence is SEQ ID NO: 112; (13) the selectedVH sequence is SEQ ID NO: 121, and the selected VL sequence is SEQ IDNO: 122; (14) the selected VH sequence is SEQ ID NO: 131, and theselected VL sequence is SEQ ID NO: 132; (15) the selected VH sequence isSEQ ID NO: 141, and the selected VL sequence is SEQ ID NO: 142; (16) theselected VH sequence is SEQ ID NO: 151, and the selected VL sequence isSEQ ID NO: 152; (17) the selected VH sequence is SEQ ID NO: 161, and theselected VL sequence is SEQ ID NO: 162; and (18) the selected VHsequence is SEQ ID NO: 171, and the selected VL sequence is SEQ ID NO:172; (19) the selected VH sequence is SEQ ID NO: 181, and the selectedVL sequence is SEQ ID NO: 182; (20) the selected VH sequence is SEQ IDNO: 191, and the selected VL sequence is SEQ ID NO: 192; (21) theselected VH sequence is SEQ ID NO: 201, and the selected VL sequence isSEQ ID NO: 202; (22) the selected VH sequence is SEQ ID NO: 211, and theselected VL sequence is SEQ ID NO: 212; (23) the selected VH sequence isSEQ ID NO: 221, and the selected VL sequence is SEQ ID NO: 222; (24) theselected VH sequence is SEQ ID NO: 231, and the selected VL sequence isSEQ ID NO: 232; (25) the selected VH sequence is SEQ ID NO: 241, and theselected VL sequence is SEQ ID NO: 242; (26) the selected VH sequence isSEQ ID NO: 251, and the selected VL sequence is SEQ ID NO: 252; (27) theselected VH sequence is SEQ ID NO: 261, and the selected VL sequence isSEQ ID NO: 262; (28) the selected VH sequence is SEQ ID NO: 271, and theselected VL sequence is SEQ ID NO: 272; (29) the selected VH sequence isSEQ ID NO: 281, and the selected VL sequence is SEQ ID NO: 282; (30) theselected VH sequence is SEQ ID NO: 291, and the selected VL sequence isSEQ ID NO: 292; (31) the selected VH sequence is SEQ ID NO: 301, and theselected VL sequence is SEQ ID NO: 302; (32) the selected VH sequence isSEQ ID NO: 311, and the selected VL sequence is SEQ ID NO: 312; (33) theselected VH sequence is SEQ ID NO: 321, and the selected VL sequence isSEQ ID NO: 322; (34) the selected VH sequence is SEQ ID NO: 331, and theselected VL sequence is SEQ ID NO: 332; (35) the selected VH sequence isSEQ ID NO: 341, and the selected VL sequence is SEQ ID NO: 342; (36) theselected VH sequence is SEQ ID NO: 351, and the selected VL sequence isSEQ ID NO: 352; (37) the selected VH sequence is SEQ ID NO: 361, and theselected VL sequence is SEQ ID NO: 362; (38) the selected VH sequence isSEQ ID NO: 371, and the selected VL sequence is SEQ ID NO: 372; (39) theselected VH sequence is SEQ ID NO: 381, and the selected VL sequence isSEQ ID NO: 382; and (40) the selected VH sequence is SEQ ID NO: 391, andthe selected VL sequence is SEQ ID NO: 392, wherein the antibody orantigen-binding fragment thereof specifically binds to CUBdomain-containing protein 1 (CDCP1).

In one embodiment the antibody or antigen-binding fragment specificallybinds to human CDCP1 and/or Cynomolgus CDCP1.

In one embodiment the antibody or antigen-binding fragment has adissociation constant (K_(d)) for human CDCP1 that is less than 10 nM,and/or a dissociation constant (K_(d)) for Cynomolgus CDCP1 that is lessthan 10 nM.

In one embodiment the antibody or antigen-binding fragment is ahumanized antibody or antigen-binding fragment thereof.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 1 and the VL comprises thesequence of SEQ ID NO: 2.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 11 and the VL comprises thesequence of SEQ ID NO: 12.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 21 and the VL comprises thesequence of SEQ ID NO: 22.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 31 and the VL comprises thesequence of SEQ ID NO: 32.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 41 and the VL comprises thesequence of SEQ ID NO: 42.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 51 and the VL comprises thesequence of SEQ ID NO: 52.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 61 and the VL comprises thesequence of SEQ ID NO: 62.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 71 and the VL comprises thesequence of SEQ ID NO: 72.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 81 and the VL comprises thesequence of SEQ ID NO: 82.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 91 and the VL comprises thesequence of SEQ ID NO: 92.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 101 and the VL comprisesthe sequence of SEQ ID NO: 102.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 111 and the VL comprisesthe sequence of SEQ ID NO: 112.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 121 and the VL comprisesthe sequence of SEQ ID NO: 122.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 131 and the VL comprisesthe sequence of SEQ ID NO: 132.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 141 and the VL comprisesthe sequence of SEQ ID NO: 142.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 151 and the VL comprisesthe sequence of SEQ ID NO:152.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 161 and the VL comprisesthe sequence of SEQ ID NO: 162.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 171 and the VL comprisesthe sequence of SEQ ID NO: 172.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO:181 1 and the VL comprisesthe sequence of SEQ ID NO: 182.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 191 and the VL comprisesthe sequence of SEQ ID NO: 192.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 201 and the VL comprisesthe sequence of SEQ ID NO: 202.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 211 and the VL comprisesthe sequence of SEQ ID NO: 212.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 221 and the VL comprisesthe sequence of SEQ ID NO: 222.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 231 and the VL comprisesthe sequence of SEQ ID NO: 232.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 241 and the VL comprisesthe sequence of SEQ ID NO: 242.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 251 and the VL comprisesthe sequence of SEQ ID NO: 252.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 261 and the VL comprisesthe sequence of SEQ ID NO: 262.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 271 and the VL comprisesthe sequence of SEQ ID NO: 272.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 281 and the VL comprisesthe sequence of SEQ ID NO: 282.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 291 and the VL comprisesthe sequence of SEQ ID NO: 292.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 301 and the VL comprisesthe sequence of SEQ ID NO: 302.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 311 and the VL comprisesthe sequence of SEQ ID NO: 312.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 321 and the VL comprisesthe sequence of SEQ ID NO: 322.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 331 and the VL comprisesthe sequence of SEQ ID NO: 332.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 341 and the VL comprisesthe sequence of SEQ ID NO: 342.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 351 and the VL comprisesthe sequence of SEQ ID NO: 352.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 361 and the VL comprisesthe sequence of SEQ ID NO: 362.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 371 and the VL comprisesthe sequence of SEQ ID NO: 372.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 381 and the VL comprisesthe sequence of SEQ ID NO: 382.

In one embodiment the VH of the antibody or antigen-binding fragmentthereof comprises the sequence of SEQ ID NO: 391 and the VL comprisesthe sequence of SEQ ID NO: 392, wherein the antibody or antigen-bindingfragment thereof specifically binds to CUB domain-containing protein 1(CDCP1).

In one embodiment, the antibody or antigen-binding fragment thereofbinds to human CDCP1, and blocks cleavage of human CDCP1 at residue 342.In one embodiment, the antibody or antigen-binding fragment thereofcomprises a VH comprising CDRs 1, 2, 3 with amino acid sequences setforth in SEQ ID NOs: 85, 86, and 87, and a VL comprising CDRs 1, 2, 3with amino acid sequences set forth in SEQ ID NOs: 88, 89, and 90. Inone embodiment, the antibody or antigen-binding fragment thereofcomprises a VH comprising CDRs 1, 2, 3 with amino acid sequences setforth in SEQ ID NOs: 95, 96, and 97, and a VL comprising CDRs 1, 2, 3with amino acid sequences set forth in SEQ ID NOs: 98, 99, and 100.

In another aspect, the disclosure relates to antibody drug conjugateshaving an antibody or antigen-binding fragment thereof disclosed herein,and a therapeutic agent.

In one embodiment, the an antibody drug conjugate comprises the antibodyor antigen-binding fragment thereof and a therapeutic agent. In oneembodiment, the therapeutic agent of the antibody drug conjugate is acytotoxic or cytostatic agent. In one embodiment, the cytotoxic orcytostatic agent in the antibody drug conjugate is a microtubuleinhibitor or a DNA alkylator. In one embodiment, the cytotoxic orcytostatic agent in the antibody drug conjugate is selected from thegroup consisting of DM4, MMAE, PDX, PDB, and IGN. In one embodiment, theagent is DM4. In another embodiment, the agent is MMAE. In oneembodiment, the agent is PDX. In one embodiment, the agent is PDB. Inone embodiment, the agent is IGN. In one embodiment, the antibody orantigen-binding fragment in the antibody drug conjugate is linked to thetherapeutic agent by a linker.

In one embodiment, the linker in the antibody drug conjugate is selectedfrom the group consisting of a cleavable peptide, a charged hindereddisulfide, and maleimido-caproyl-valine-citrulline.

In one embodiment, the therapeutic agent in the antibody drug conjugateis DM4, and the linker is D-Ala-L-Ala dpa.

In one embodiment, the therapeutic agent in the antibody drug conjugateis DM4, and the linker is N-succinimidyl4-(2-pyridyldithio)-2-sulfobutanoate (sSPDB).

In one embodiment, the therapeutic agent in the antibody drug conjugateis MMAE, and the linker is maleimido-caproyl-valine-citrulline (MC-VC).

In one embodiment, therapeutic agent in the antibody drug conjugate isIGN, and the linker is D-Ala-L-Ala dpa.

In one aspect the present invention provides a an antibody drugconjugate has the formula Ab-[L-D]n, wherein Ab comprises the antibodyor antigen-binding fragment thereof wherein L comprises an optionallinker; D is a therapeutic agent; and n is an integer from about 1 toabout 20.

In one embodiment, a method of treating cancer in a subject is provided,comprising: identifying a subject having cancer; and administering tothe subject a pharmaceutical composition comprising a therapeuticallyeffective amount of the antibody drug conjugate.

In one embodiment, the cancer is breast cancer, lung cancer, small celllung cancer, liver cancer, pancreatic cancer, ovarian cancer, kidneycancer or colon cancer.

In one embodiment, the cancer is triple negative breast cancer. In oneembodiment, the therapeutic agent in the antibody drug conjugate is DM4,and the linker is D-Ala-L-Ala dpa or sSPDB. In one embodiment, thetherapeutic agent in the antibody drug conjugate is MMAE, and the linkeris MC-VC.

In one embodiment, the cancer is colon cancer. In one embodiment, thetherapeutic agent in the antibody drug conjugate is IGN, and the linkeris D-Ala-L-Ala dpa. In one embodiment, the therapeutic agent in theantibody drug conjugate is DM4, and the linker is D-Ala-L-Ala dpa orsSPDB. In one embodiment, the therapeutic agent in the antibody drugconjugate is MMAE, and the linker is MC-VC.

In one embodiment, the cancer is small cell lung cancer. In oneembodiment, the therapeutic agent in the antibody drug conjugate is IGN,and the linker is D-Ala-L-Ala dpa.

In another aspect, the present invention provides a method of treatingcancer in a subject is provided, comprising identifying a subject havingcancer; and administering to the subject a pharmaceutical compositioncomprising a therapeutically effective amount of the antibody orantigen-binding fragment thereof, or a therapeutically effective amountof the antibody drug conjugate.

In one embodiment, the cancer is breast cancer, triple negative breastcancer, lung cancer, small cell lung cancer, liver cancer, pancreaticcancer, ovarian cancer, kidney cancer or colon cancer.

In another aspect, the disclosure relates to a polynucleotide linked toa heterologous nucleic acid, wherein the polynucleotide is selected fromthe group consisting of (a) a polynucleotide encoding an immunoglobulinheavy chain or a fragment thereof comprising a heavy chain variableregion (VH) comprising complementarity determining regions (CDRs) 1, 2,and 3 with the amino acid sequences set forth in FIG. 20 , and whereinthe VH when paired with the corresponding light chain variable region(VL) as shown in FIG. 15 binds to CDCP1 protein; (b) a polynucleotideencoding an immunoglobulin light chain or a fragment thereof comprisinga VL comprising CDRs 1, 2, and 3 with the amino acid sequences set forthin FIG. 20 , and wherein the VL when paired with the corresponding VH asshown in FIG. 15 binds to CDCP1; (c) a polynucleotide encoding (i) animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inFIG. 20 ; and (ii) an immunoglobulin light chain or a fragment thereofcomprising a VL comprising CDRs 1, 2, and 3 with the amino acidsequences set forth in FIG. 20 ; and (d) a polynucleotide encoding animmunoglobulin heavy chain or a fragment thereof comprising a VHcomprising the amino acid sequence set forth in FIG. 18 , wherein the VHwhen paired with the corresponding VL as shown in FIG. 15 binds toCDCP1.

In one aspect, the present invention an expression vector comprising aheterologous promoter operably linked to a polynucleotide encoding apolypeptide disclosed herein.

In another aspect, the disclosure provides an expression vectorcomprising: a first polynucleotide encoding a first polypeptidecomprising an immunoglobulin heavy chain or a fragment thereofcomprising a heavy chain variable region (VH) comprising VHcomplementarity determining regions (CDRs) 1, 2, and 3 with the aminoacid sequences set forth in FIG. 20 ; and a second polynucleotideencoding a second polypeptide comprising an immunoglobulin light chainor a fragment thereof comprising a light chain variable region (VL)comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forthin FIG. 20 , wherein the immunoglobulin heavy chain or fragment thereofwhen paired with the immunoglobulin light chain or fragment thereofforms an anti-CDCP1 antibody or CDCP1-binding fragment thereof, andwherein the expression vector is, for example, a plasmid, phage, orvirus.

In some embodiments, the complementarity determining region (CDR)sequences in the heavy chain variable region and the light chainvariable region comprise or consist of the CDR sequences as set forth inFIG. 20 . In some embodiments, the CDR sequences in the heavy chainvariable region and the light chain variable region are at least 80%,90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to the CDR sequences asset forth in FIG. 20 . In some embodiments, the CDR sequences in theheavy chain variable region and the light chain variable region differfrom the CDR sequences in FIG. 20 by one, two, three, four, or fiveamino acids. In some embodiments, the CDRs in FIG. 20 are substituted byone, two, three, four, or five conservative amino acids.

In another aspect, the present invention provides compositions thatinclude at least one antibody disclosed herein conjugated to atherapeutic agent, e.g., a cytotoxic drug or radioisotope, and/or to areporter group. In some implementations, an antibody that is conjugatedto a therapeutic agent is administered to a subject to induce cell deathof a cancer cell, e.g., a cancer cell that expresses or overexpressesCDCP1 protein, or has CDCP1 protein on the cell surface.

In some embodiments, the antibodies are conjugated to a detectablemarker. The conjugates are administered to a subject to detect a cancercell that expresses CDCP1 protein and/or has a detectable and/orelevated level of a CDCP1 protein.

In some embodiments, an antibody is linked (e.g., covalently bonded,hydrogen bonded, or ionicly bonded) to a surface (e.g., a microfluidicdevice, a chromatography resin, an array, polymer, or a bead).

In a further aspect, the present invention provides compositions thatinclude at least one of the antibodies disclosed herein and apharmaceutically acceptable excipient. In some embodiments, thedisclosure features dry (e.g., lyophilized) compositions that includeone or more antibodies disclosed herein and, optionally, one or morepharmaceutically acceptable excipients.

In another aspect, the present invention provides polynucleotides, e.g.,DNA, that encode a polypeptide chain, e.g., an antibody heavy or lightchain, of any of the antibodies disclosed herein. For example, thepolynucleotide may include a sequence disclosed herein. In someimplementations, the polynucleotides, e.g., DNA, do not include introns.The disclosure also features vectors, e.g., recombinant vectors andexpression vectors that include the above polynucleotides, and a cell,e.g., an isolated cell, e.g., recombinant cells or hybridomas, thatinclude the above polynucleotides and/or vectors. In someimplementations, the vector is stably integrated into a chromosome ofthe cell, e.g., a mammalian cell, bacterial cell, or yeast cell. In someimplementations, the disclosure features methods of producing antibodiesthat include culturing the cells, e.g., isolated cells, under conditionswhere the antibodies are expressed, and methods of collecting theantibodies.

The present invention also provides the antibodies, nucleic acids,compositions, and cells disclosed herein and the use thereof fortreatment, prophylaxis, imaging, and/or diagnosis of a cancer.

In some embodiments, the composition is formulated for intravenousadministration. In some implementations, the cancer expresses (e.g.,overexpresses) CDCP1 or has CDCP1 on the cell surface. In someimplementations, the cancer is characterized by the presence of a CDCP1and/or an elevated level of a CDCP1 protein (e.g., as compared to areference level, e.g., a level of a CDCP1 protein in a CDCP1 proteinproduced by a healthy subject) produced by the cancer cells.

In a further aspect, the present invention provides methods fortreatment of a cancer (e.g., a cancer characterized by overexpression ofCDCP1 in cancer cells, or a cancer characterized by having CDCP1 on thesurface of the cancer cells) (e.g., breast cancer, triple-negativebreast cancer, carcinoid cancer, cervical cancer, endometrial cancer,glioma, head and neck cancer, liver cancer, lung cancer, small cell lungcancer, lymphoma, melanoma, ovarian cancer, pancreatic cancer, prostatecancer, renal cancer, gastric cancer, testicular cancer, thyroid cancer,bladder cancer, urethral cancer, colorectal cancer, or hematologicmalignancy), that include administering an antibody, nucleic acid,composition, or cell disclosed herein to a subject with a cancer in atherapeutically effective amount.

In another aspect, the present invention provides methods that includeadministering an antibody or composition, e.g., a cell composition,antibody-drug conjugate, or antibody-radioisotope conjugate disclosedherein to a subject in need thereof, e.g., a subject having, oridentified or diagnosed as having a cancer characterized byoverexpression of CDCP1 in cancer cells, or a cancer characterized byhaving CDCP1 on the surface of the cancer cells, e.g., breast cancer,triple-negative breast cancer, carcinoid cancer, cervical cancer,endometrial cancer, glioma, head and neck cancer, liver cancer, lungcancer, small cell lung cancer, non-small cell lung cancer, lymphoma,melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renalcancer, gastric cancer, testicular cancer, thyroid cancer, bladdercancer, urethral cancer, colorectal cancer, colon cancer, or hematologicmalignancy cells.

In some embodiments, the subject is identified as being a subject whoexpresses CDCP1, e.g., using any of the methods described herein, or hasan elevated level of a CDCP1 protein, e.g., as compared to a referencelevel, e.g., a level of a CDCP1 protein in a CDCP1 protein produced by ahealthy subject, a level of a CDCP1 protein in CDCP1 protein produced bya non-cancerous, e.g., primary cell, or a threshold level of a CDCP1protein, in which a determined level of a CDCP1 protein that is abovethis value indicates that the subject should be administered an antibodydescribed herein.

In yet another aspect, the present invention provides methods for cancerprophylaxis (or reducing a subject's risk of developing a cancercharacterized by expression, e.g., overexpression, of CDCP1 protein incancer cells or a cancer characterized by having CDCP1 on the surface ofthe cancer cells, e.g., breast cancer (e.g., triple-negative breastcancer), carcinoid cancer, cervical cancer, endometrial cancer, glioma,head and neck cancer, liver cancer, lung cancer, small cell lung cancer,lymphoma, melanoma, ovarian cancer, pancreatic cancer, prostate cancer,renal cancer, gastric cancer, testicular cancer, thyroid cancer, bladdercancer, urethral cancer, colorectal cancer, or hematologic malignancycells (e.g., as compared to a subject at risk for developing cancer butreceiving no treatment or receiving a different treatment). The methodsinclude administering an antibody, nucleic acid, composition, or celldisclosed herein to a subject in need thereof in a prophylacticallyeffective amount.

In some embodiments, the cancer expresses CDCP1 protein. In someembodiments, the cancer cells have CDCP1 on the cell surface. In someimplementations of any of these methods, the subject is identified ashaving an elevated risk of developing cancer.

In another aspect, the present invention provides methods of detecting aCDCP1 protein (e.g., a CDCP1 protein) in a sample (e.g., a biopsysample). The methods include contacting a sample with an antibodydisclosed herein and detecting binding of the agent to the sample,thereby detecting CDCP1 protein in the sample. Some implementationsfurther include recording the detection or non-detection of CDCP1protein in the clinical records of a subject from whom the sample wasobtained. In some implementations, the clinical record is stored on atangible computer readable medium, e.g., a disc, magnetic tape, orcomputer memory.

In some embodiments of any of the methods described herein, theantibodies described herein are contacted with a sample and/or a cell,and the antibody to CDCP1 can be used in an immunoassay (e.g., anenzyme-linked immunosorbent assay), fluorescence-assisted cell sorting,microfluidics, and chromatography.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a schematic diagram showing CDCP1 constructs expressing fulllength CDCP1 (FL-CDCP1) or cleaved CDCP1 (Clv-CDCP1(N342)) (having aminoacids 343-836).

FIG. 1B are graphs showing flow cytometry analysis of two CDCP1antibodies on live 293T, 293T/FL-CDCP1, and 293T/Clv N342 cells.

FIG. 1C is an image of a Western blot of lysates of 293T cells stablytransfected with FL or two forms of Clv-CDCP1: 8368 (AA369-836), andN342: (AA343-836).

FIG. 2A is a schematic diagram showing a protocol for a R368 cleavageblocking assay. This Figure discloses “8×His” as SEQ ID NO: 406.

FIGS. 2B-2C are images of Western blots showing the results of a R368cleavage blocking assay.

FIG. 3 is a graph showing that 18C6 antibody binds to cleaved CDCP(AA343-836) (Clv-CDCP1(N342)) at a higher affinity than to full lengthCDCP1.

FIG. 4 is a schematic diagram showing distinct epitope bins for 18selected recombinant human antibodies.

FIGS. 5A-5E are graphs showing the in vitro killing effect of antibodyvcMMAE conjugates on cells from BrCa cell lines MCF7, BT549, MDA231, andcells from CRC cell lines SW48 and HCT116, wherein the antibodies in theantibody vcMMAE conjugates are hIgG1, 47G7, 41A09, 38E11, 27H10, 18C6,and 03B11.

FIGS. 6A-6C are graphs showing antibody dependent cytotoxicity (ADCC)effects of hIgG (control) and human CDCP1 antibodies 2F3, 10E2, 38E11,and 41A9 on cells from TNBC cell lines BT549 and MDA231, and cells fromCRC cell line SW48.

FIGS. 7A-7G are graphs showing complement-dependent cytotoxicity (CDC)effect of hIgG (control) and human CDCP1 antibodies 27H10, 38E11, 3B11,47G7, 41A9 and 18C6 on BT549 cells.

FIG. 8A is a graph showing the antibody vcMMAE conjugate (41A10-vcMMAE)inhibited tumor growth in mice xenografted with cells from TNBC cellline MDA231 in vivo.

FIG. 8B is a graph showing body weight plot for mice with MDA231 tumortreated with the antibody vcMMAE conjugate (41A10-vcMMAE).

FIG. 9 is a graph showing human CDCP1 antibodies conjugated with vcMMAEinhibited SW48 tumor growth in mice in vivo, wherein the antibodies inthe antibody vcMMAE conjugates are hIgG1 (control), 47G7, 41A09, 38E11,27H10, 18C6, and 03B11.

FIG. 10 is a graph showing that human CDCP1 antibodies conjugated withvcMMAE inhibited tumor growth in mice xenografted with cells from TNBCcell line MDA231 in vivo, wherein the antibodies in the conjugates are18C6, 27H10, 38E11, 41A9, and hIgG1.

FIG. 11 is a graph showing that human CDCP1 antibodies conjugated withvcMMAE caused regression of established SW48 tumors in mice in vivo,wherein the antibodies in the conjugates are 18C6, 27H10, 38E11, 41A9,and hIgG1.

FIG. 12 is a graph showing the survival plot for mice with SW48 tumorstreated with the antibody vcMMAE conjugates as shown in FIG. 11 .

FIG. 13A is a graph showing dose-dependent tumor growth inhibitoryeffects of 38E11-MMAE on SW48 tumors in mice.

FIG. 13B is a graph showing dose-dependent tumor growth inhibitoryeffects of 271410-MMAE on SW48 tumors in mice.

FIG. 13C is a graph showing dose-dependent tumor growth inhibitoryeffects of 18C6-MMAE on SW48 tumors in mice.

FIG. 13D is a graph showing dose-dependent tumor growth inhibitoryeffects of 10E2-MMAE on SW48 tumors in mice.

FIG. 14A is a graph showing Kaplan Meier analysis of SW48 tumor bearingmice treated with PBS, hIgG1-MMAE, or 38E11-MMAE at 0.5, 1.5, 5 mg/kg.

FIG. 14B is a graph showing Kaplan Meier analysis of SW48 tumor bearingmice treated with PBS, hIgG1-MMAE, or 27H10-MMAE at 0.5, 1.5, 5 mg/kg.

FIG. 14C is a graph showing Kaplan Meier analysis of SW48 tumor bearingmice treated with PBS, hIgG1-MMAE, or 18C6-MMAE at 0.5, 1.5, 5 mg/kg.

FIG. 14D is a graph showing Kaplan Meier analysis of SW48 tumor bearingmice treated with PBS, hIgG1-MMAE, or 10E2-MMAE at 0.5, 1.5, 5 mg/kg.

FIG. 15 lists the SEQ ID numbers for the nucleic acid sequences of thevariable regions of the heavy chain and the light chain, and the aminoacid sequences of the variable regions and complementarity determiningregions (CDRs) of the heavy chain and the light chain of 40 anti-CDCP1antibodies.

FIG. 16 shows the properties of 40 anti-CDCP1 antibodies.

FIG. 17 shows the properties of 18 anti-CDCP1 antibodies, which are asubset of the 40 anti-CDCP1 antibodies in FIG. 16 .

FIG. 18 lists the amino acid sequence of the heavy chain variable region(VH) and the light chain variable region (VL) of the 40 anti-CDCP1antibodies in FIG. 15 .

FIG. 19 lists the nucleotide sequence of the heavy chain variable regionand the light chain variable region of the 40 anti-CDCP1 antibodies inFIG. 15 .

FIG. 20 lists the amino acid sequence of the complementary determiningregions in the heavy chain variable region (VH CDR1, VH CDR2, VH CDR3)and complementary determining regions in the light chain variable region(VL CDR1, VL CDR2, VL CDR3)) of the 40 anti-CDCP1 antibodies in FIG. 15.

FIGS. 21A and 21B are graphs showing the in vitro killing effect ofhIgG1-vcMMAE and 38E11-vcMMAE on prostate cancer cells.

FIGS. 22A and 22B are graphs showing the in vitro killing effect ofhIgG1-PBD and 38E11-PBD on prostate cancer cells.

FIGS. 23A and 23B are graphs showing the in vitro killing effect ofhIgG1-vcMMAE and 38E11-vcMMAE on NSLC cells.

FIGS. 24A and 24B are graphs showing the in vitro killing effect ofhIgG1-PBD and 38E11-PBD on NSLC cells.

FIGS. 25A and 25B are graphs showing the tumor growth of eight TNBC PDXmodels in response to 38E11-vcMMAE and hIgG-vcMMAE.

FIGS. 26A and 26B are graphs showing the body weight plots of eight TNBCPDX models in response to 38E11-vcMMAE and hIgG-vcMMAE.

DETAILED DESCRIPTION

Various aspects of the disclosure relate to anti-CUB domain-containingprotein 1 (anti-CDCP1) antibodies and antibody fragments, anti-CDCP1ADCs, and pharmaceutical compositions thereof, as well as nucleic acids,recombinant expression vectors and host cells for making such antibodiesand fragments. Methods of using the antibodies and ADCs described hereinto detect human CDCP1, to bind to and inhibit human CDCP1 on CDCP1expressing cells, to inhibit CDCP1 signaling, in vivo, and/or to treatCDCP1-associated disorders, e.g., cancer, including, but not limited to,breast cancer, e.g., triple negative breast cancer, lung cancer, e.g.,non-small cell lung cancer (NSCLC), liver cancer, pancreatic cancer,ovarian cancer, kidney cancer, and colon cancer. In one embodiment, theanti-CDCP1 antibodies or ADCs of the invention are administered incombination with one or more immune checkpoint inhibitors (e.g.,antibody or small molecule immune checkpoint inhibitors) for thetreatment of a cancer. In another embodiment of the invention,anti-CDCP1 antibody drug conjugates (ADCs) of the invention (e.g., theCDCP1 antibodies of the invention conjugated to a toxin) areinternalized and induce cell death of cells endogenously expressingCDCP1.

In one embodiment, the anti-CDCP1 antibodies, bispecific antibodies, orADCs disclosed herein are administered in combination with a PARP (polyADP ribose polymerase) inhibitor. PARP inhibitors are well known tothose of ordinary skill in the art and include, but are not limited to,Niraparib, Olaparib, Rucaparib, Iniparib, Talazoparib, Veliparib, CEP9722, E7016, BGB-290, and 3-aminobenazamine.

I. Definitions

In order that the invention may be more readily understood, certainterms are first defined. In addition, it should be noted that whenever avalue or range of values of a parameter are recited, it is intended thatvalues and ranges intermediate to the recited values are also intendedto be part of this invention.

The terms “CUB domain-containing protein 1 antibody” or “anti-CDCP1antibody”, used interchangeably herein, refer to an antibody thatspecifically binds to CDCP1, e.g., human CDCP1. An antibody “whichbinds” an antigen of interest, i.e., CDCP1, is one capable of bindingthat antigen with sufficient affinity such that the antibody is usefulin targeting a cell expressing the antigen. In a preferred embodiment,the antibody specifically binds to human CDCP1 (hCDCP1). Examples ofanti-CDCP1 antibodies are disclosed in the Examples, below. Unlessotherwise indicated, the term “anti-CDCP1 antibody” is meant to refer toan antibody which binds to wild type CDCP1, a variant, or an isoform ofCDCP1.

Alternative splicing results in at least two transcript variants ofhCDCP1. CDCP1 nucleotide and polypeptide sequences are reported asAccession Nos. NM_022842.4 (transcript variant 1 mRNA), NP_073753.3(isoform 1 polypeptide) and NM_178181.2 (transcript variant 2 mRNA),NP_835488.1 (isoform 2 polypeptide). CDCP1 contains a CUB domain atpositions 225-297 of NP_073753.3. The extracellular domain of theprotein described in NP_073753.3 includes amino acid residues 30-667.

The terms “specific binding” or “specifically binding”, as used herein,in reference to the interaction of a CDCP1 antibody or an ADC with asecond chemical species, mean that the interaction is dependent upon thepresence of a particular structure (e.g., an antigenic determinant orepitope) on the chemical species; for example, an antibody recognizesand binds to a specific protein structure rather than to proteinsgenerally. If an antibody or ADC is specific for epitope “A”, thepresence of a molecule containing epitope A (or free, unlabeled A), in areaction containing labeled “A” and the antibody, will reduce the amountof labeled A bound to the antibody or ADC.

In one embodiment, the phrase “specifically binds to hCDCP1” or“specific binding to hCDCP1”, as used herein, refers to the ability ofan anti-CDCP1 antibody or ADC to interact with CDCP1 (human orcynomolgus monkey CDCP1) with a dissociation constant (K_(D)) of about2,000 nM or less, about 1,000 nM or less, about 500 nM or less, about200 nM or less, about 100 nM or less, about 75 nM or less, about 25 nMor less, about 21 nM or less, about 12 nM or less, about 11 nM or less,about 10 nM or less, about 9 nM or less, about 8 nM or less, about 7 nMor less, about 6 nM or less, about 5 nM or less, about 4 nM or less,about 3 nM or less, about 2 nM or less, about 1 nM or less, about 0.5 nMor less, about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM orless, or about 0.001 nM or less. In another embodiment, the phrase“specifically binds to hCDCP1” or “specific binding to hCDCP1”, as usedherein, refers to the ability of an anti-CDCP1 antibody or ADC tointeract with hCDCP1 with a dissociation constant (K_(D)) of betweenabout 1 μM (0.001 nM) to 2,000 nM, between about 500 μM (0.5 nM) to1,000 nM, between about 500 μM (0.5 nM) to 500 nM, between about 1 nM)to 200 nM, between about 1 nM to 100 nM, between about 1 nM to 50 nM,between about 1 nM to 20 nM, or between about 1 nM to 5 nM. In oneembodiment, K_(D) is determined by surface plasmon resonance orBio-Layer Interferometry, or by any other method known in the art.Bio-Layer Interferometry refers to an optical phenomenon that allows forthe analysis of real-time biospecific interactions by measuring theinterference patterns of reflected white light, for example using theOctet™ system (ForteBio, Pall Corp. Fremont, Calif.). For furtherdescription of the Octet™ system, see Li, B et al. (2011) J. Pharm.Blamed. Anal. 54(2):286-294 and Abdiche, Y. N., et al. (2009) Anal.Biochem. 386(2):172-180, the contents of which are incorporated hereinby reference.

The term “antibody” broadly refers to an immunoglobulin (Ig) molecule,generally comprised of four polypeptide chains, two heavy (H) chains andtwo light (L) chains, or any functional fragment, mutant, variant, orderivative thereof, that retains the essential target binding featuresof an Ig molecule. Such mutant, variant, or derivative antibody formatsare known in the art. Non-limiting embodiments of which are discussedbelow.

In a full-length antibody, each heavy chain is comprised of a heavychain variable region (abbreviated herein as HCVR or VH) and a heavychain constant region. The heavy chain constant region is comprised ofthree domains, CH1, CH2 and CH3. Each light chain is comprised of alight chain variable region (abbreviated herein as LCVR or VL) and alight chain constant region. The light chain constant region iscomprised of one domain, CL. The VH and VL regions can be furthersubdivided into regions of hypervariability, termed complementaritydetermining regions (CDR), interspersed with regions that are moreconserved, termed framework regions (FR). Each VH and VL is composed ofthree CDRs and four FRs, arranged from amino-terminus tocarboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4. Immunoglobulin molecules can be of any type (e.g., IgG, IgE,IgM, IgD, IgA and IgY) and class (e.g., IgG1, IgG2, IgG 3, IgG4, IgA1and IgA2) or subclass.

The term “antigen binding portion” of an antibody (or simply “antibodyportion”), as used herein, refers to one or more fragments of anantibody that retain the ability to specifically bind to an antigen(e.g., hCDCP1). It has been shown that the antigen binding function ofan antibody can be performed by fragments of a full-length antibody.Such antibody embodiments may also be bispecific, dual specific, ormulti-specific formats; specifically binding to two or more differentantigens. Examples of binding fragments encompassed within the term“antigen binding portion” of an antibody include (i) a Fab fragment, amonovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) aF(ab′)₂ fragment, a bivalent fragment comprising two Fab fragmentslinked by a disulfide bridge at the hinge region; (iii) a Fd fragmentconsisting of the VH and CH1 domains; (iv) a Fv fragment consisting ofthe VL and VH domains of a single arm of an antibody, (v) a dAb fragment(Ward et al., (1989) Nature 341:544-546, Winter et al., PCT publicationWO 90/05144 A1 herein incorporated by reference), which comprises asingle variable domain; and (vi) an isolated complementarity determiningregion (CDR). Furthermore, although the two domains of the Fv fragment,VL and VH, are coded for by separate genes, they can be joined, usingrecombinant methods, by a synthetic linker that enables them to be madeas a single protein chain in which the VL and VH regions pair to formmonovalent molecules (known as single chain Fv (scFv); see e.g., Bird etal. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl.Acad. Sci. USA 85:5879-5883). Such single chain antibodies are alsointended to be encompassed within the term “antigen binding portion” ofan antibody. In certain embodiments, scFv molecules may be incorporatedinto a fusion protein. Other forms of single chain antibodies, such asdiabodies are also encompassed. Diabodies are bivalent, bispecificantibodies in which VH and VL domains are expressed on a singlepolypeptide chain, but using a linker that is too short to allow forpairing between the two domains on the same chain, thereby forcing thedomains to pair with complementary domains of another chain and creatingtwo antigen binding sites (see e.g., Holliger, P., et al. (1993) Proc.Natl. Acad. Sci. USA 90:6444-6448; Poljak, R. J., et al. (1994)Structure 2:1121-1123). Such antibody binding portions are known in theart (Kontermann and Dubel eds., Antibody Engineering (2001)Springer-Verlag. New York. 790 pp. (ISBN 3-540-41354-5).

The term “antibody construct” as used herein refers to a polypeptidecomprising one or more the antigen binding portions disclosed hereinlinked to a linker polypeptide or an immunoglobulin constant domain.Linker polypeptides comprise two or more amino acid residues joined bypeptide bonds and are used to link one or more antigen binding portions.Such linker polypeptides are well known in the art (see e.g., Holliger,P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R.J., et al. (1994) Structure 2:1121-1123). An immunoglobulin constantdomain refers to a heavy or light chain constant domain. Antibodyportions, such as Fab and F(ab′)₂ fragments, can be prepared from wholeantibodies using conventional techniques, such as papain or pepsindigestion, respectively, of whole antibodies. Moreover, antibodies,antibody portions and immunoadhesion molecules can be obtained usingstandard recombinant DNA techniques, as described herein.

An “isolated antibody”, as used herein, is intended to refer to anantibody that is substantially free of other antibodies having differentantigenic specificities (e.g., an isolated antibody that specificallybinds CDCP1 is substantially free of antibodies that specifically bindantigens other than CDCP1). An isolated antibody that specifically bindsCDCP1 may, however, have cross-reactivity to other antigens, such asCDCP1 molecules from other species. Moreover, an isolated antibody maybe substantially free of other cellular material and/or chemicals.

The term “humanized antibody” refers to antibodies which comprise heavyand light chain variable region sequences from a nonhuman species (e.g.,a mouse) but in which at least a portion of the VH and/or VL sequencehas been altered to be more “human-like”, i.e., more similar to humangermline variable sequences. In particular, the term “humanizedantibody” is an antibody or a variant, derivative, analog or fragmentthereof which immunospecifically binds to an antigen of interest andwhich comprises a framework (FR) region having substantially the aminoacid sequence of a human antibody and a complementary determining region(CDR) having substantially the amino acid sequence of a non-humanantibody. As used herein, the term “substantially” in the context of aCDR refers to a CDR having an amino acid sequence at least 80%,preferably at least 85%, at least 90%, at least 95%, at least 98% or atleast 99% identical to the amino acid sequence of a non-human antibodyCDR. A humanized antibody comprises substantially all of at least one,and typically two, variable domains (Fab, Fab′, F(ab′)₂, FabC, Fv) inwhich all or substantially all of the CDR regions correspond to those ofa non-human immunoglobulin (i.e., donor antibody) and all orsubstantially all of the framework regions are those of a humanimmunoglobulin consensus sequence. Preferably, a humanized antibody alsocomprises at least a portion of an immunoglobulin constant region (Fe),typically that of a human immunoglobulin. In some embodiments, ahumanized antibody contains both the light chain as well as at least thevariable domain of a heavy chain. The antibody also may include the CH1,hinge, CH2, CH3, and CH4 regions of the heavy chain. In someembodiments, a humanized antibody only contains a humanized light chain.In other embodiments, a humanized antibody only contains a humanizedheavy chain. In specific embodiments, a humanized antibody only containsa humanized variable domain of a light chain and/or humanized heavychain.

The humanized antibody can be selected from any class ofimmunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any isotype,including without limitation IgG1, IgG2, IgG3 and IgG4. The humanizedantibody may comprise sequences from more than one class or isotype, andparticular constant domains may be selected to optimize desired effectorfunctions using techniques well-known in the art.

The terms “Kabat numbering,” “Kabat definitions,” and “Kabat labeling”are used interchangeably herein. These terms, which are recognized inthe art, refer to a system of numbering amino acid residues which aremore variable (i.e., hypervariable) than other amino acid residues inthe heavy and light chain variable regions of an antibody, or an antigenbinding portion thereof (Kabat et al. (1971) Ann. NY Acad, Sci.190:382-391 and, Kabat, E. A., et al. (1991) Sequences of Proteins ofImmunological Interest, Fifth Edition, U.S. Department of Health andHuman Services, NIH Publication No. 91-3242). For the heavy chainvariable region, the hypervariable region ranges from amino acidpositions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, andamino acid positions 95 to 102 for CDR3. For the light chain variableregion, the hypervariable region ranges from amino acid positions 24 to34 for CDR1, amino acid positions 50 to 56 for CDR2, and amino acidpositions 89 to 97 for CDR3.

As used herein, the term “CDR” refers to the complementarity determiningregion within antibody variable sequences. There are three CDRs in eachof the variable regions of the heavy chain (HC) and the light chain(LC), which are designated CDR1, CDR2 and CDR3 (or specifically HC CDR1,HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3), for each of thevariable regions. The term “CDR set” as used herein refers to a group ofthree CDRs that occur in a single variable region capable of binding theantigen. The exact boundaries of these CDRs have been defineddifferently according to different systems. The system described byKabat (Kabat et at, Sequences of Proteins of Immunological Interest(National Institutes of Health, Bethesda, Md. (1987) and (1991)) notonly provides an unambiguous residue numbering system applicable to anyvariable region of an antibody, but also provides precise residueboundaries defining the three CDRs. These CDRs may be referred to asKabat CDRs. Chothia and coworkers (Chothia &Lcsk, J. Mol. Biol.196:901-917 (1987) and Chothia et al., Nature 342:877-883 (1989)) foundthat certain sub-portions within Kabat CDRs adopt nearly identicalpeptide backbone conformations, despite having great diversity at thelevel of amino acid sequence. These sub-portions were designated as L1,L2 and L3 or H1, H2 and H3 where the “L” and the “H” designates thelight chain and the heavy chains regions, respectively. These regionsmay be referred to as Chothia CDRs, which have boundaries that overlapwith Kabat CDRs. Other boundaries defining CDRs overlapping with theKabat CDRs have been described by Padlan (FASEB J. 9:133-139 (1995)) andMacCallum (J Mol Biol 262(5):732-45 (1996)). Still other CDR boundarydefinitions may not strictly follow one of the above systems, but willnonetheless overlap with the Kabat CDRs, although they may be shortenedor lengthened in light of prediction or experimental findings thatparticular residues or groups of residues or even entire CDRs do notsignificantly impact antigen binding. The methods used herein mayutilize CDRs defined according to any of these systems, althoughpreferred embodiments use Kabat or Chothia defined CDRs.

As used herein, the term “framework” or “framework sequence” refers tothe remaining sequences of a variable region minus the CDRs. Because theexact definition of a CDR sequence can be determined by differentsystems, the meaning of a framework sequence is subject tocorrespondingly different interpretations. The six CDRs (CDR-L1, CDR-L2,and CDR-L3 of light chain and CDR-H1, CDR-H2, and CDR-H3 of heavy chain)also divide the framework regions on the light chain and the heavy chaininto four sub-regions (FR1, FR2, FR3 and FR4) on each chain, in whichCDR1 is positioned between FR1 and FR2, CDR2 between FR2 and FR3, andCDR3 between FR3 and FR4. Without specifying the particular sub-regionsas FR1, FR2, FR3 or FR4, a framework region, as referred by others,represents the combined FR's within the variable region of a single,naturally occurring immunoglobulin chain. As used herein, a FRrepresents one of the four sub-regions, and FRs represents two or moreof the four sub-regions constituting a framework region.

The framework and CDR regions of a humanized antibody need notcorrespond precisely to the parental sequences, e.g., the donor antibodyCDR or the consensus framework may be mutagenized by substitution,insertion and/or deletion of at least one amino acid residue so that theCDR or framework residue at that site does not correspond to either thedonor antibody or the consensus framework. In a preferred embodiment,such mutations, however, will not be extensive. Usually, at least 80%,preferably at least 85%, more preferably at least 90%, and mostpreferably at least 95% of the humanized antibody residues willcorrespond to those of the parental FR and CDR sequences. As usedherein, the term “consensus framework” refers to the framework region inthe consensus immunoglobulin sequence. As used herein, the term“consensus immunoglobulin sequence” refers to the sequence formed fromthe most frequently occurring amino acids (or nucleotides) in a familyof related immunoglobulin sequences (See e.g., Winnaker, From Genes toClones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family ofimmunoglobulins, each position in the consensus sequence is occupied bythe amino acid occurring most frequently at that position in the family.If two amino acids occur equally frequently, either can be included inthe consensus sequence.

“Percent (%) amino acid sequence identity” with respect to a peptide orpolypeptide sequence is defined as the percentage of amino acid residuesin a candidate sequence that are identical with the amino acid residuesin the specific peptide or polypeptide sequence, after aligning thesequences and introducing gaps, if necessary, to achieve the maximumpercent sequence identity, and not considering any conservativesubstitutions as part of the sequence identity. Alignment for purposesof determining percent amino acid sequence identity can be achieved invarious ways that are within the skill in the art, for instance, usingpublicly available computer software such as BLAST, BLAST-2, ALIGN orMegalign (DNASTAR) software. Those skilled in the art can determineappropriate parameters for measuring alignment, including any algorithmsneeded to achieve maximal alignment over the full length of thesequences being compared. In one embodiment, the disclosure includes anamino acid sequence having at least 80%, at least 85%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98%, or at least 99%identity to an amino acid sequence set forth in any one of SEQ ID NOs: 1to 400 and 402-505.

The term “multivalent antibody” is used herein to denote an antibodycomprising two or more antigen binding sites. In certain embodiments,the multivalent antibody may be engineered to have the three or moreantigen binding sites, and is generally not a naturally occurringantibody.

The term “multispecific antibody” refers to an antibody capable ofbinding two or more unrelated antigens.

The term “dual variable domain” or “DVD,” as used interchangeablyherein, are antigen binding proteins that comprise two or more antigenbinding sites and are tetravalent or multivalent binding proteins. SuchDVDs may be monospecific, i.e., capable of binding one antigen ormultispecific, i.e. capable of binding two or more antigens. DVD bindingproteins comprising two heavy chain DVD polypeptides and two light chainDVD polypcptides are referred to a DVD Ig. Each half of a DVD Igcomprises a heavy chain DVD polypeptide, and a light chain DVDpolypeptide, and two antigen binding sites. Each binding site comprisesa heavy chain variable domain and a light chain variable domain with atotal of 6 CDRs involved in antigen binding per antigen binding site. Inone embodiment, the CDRs described herein are used in an anti-CDCP1 DVD.

The term “activity” includes activities such as the bindingspecificity/affinity of an antibody or ADC for an antigen, for example,an anti-hCDCP1 antibody or ADC that binds to a CDCP1 antigen. In oneembodiment, an anti-CDCP1 antibody or anti-CDCP1 ADC activity includes,but it not limited to, binding to CDCP1 in vitro; binding to CDCP1 oncells expressing CDCP1 in vivo; modulating (e.g., inhibiting) src and/orEGFR signaling; inducing cell death in cells expressing CDCP1, includingbreast cancer cells, e.g., triple negative breast cancer cells, coloncancer cells, lung cancer cells, e.g., non-small cell lung carcinoma(NSCLC) cells, liver cancer cells, pancreatic cancer cells, ovariancancer cells, kidney cancer cells; inhibiting cancer cell invasion andmetastasis; decreasing or inhibiting cancer, e.g., breast cancer, e.g.,triple negative breast cancer, colon cancer, lung cancer, e.g.,non-small cell lung carcinoma (NSCLC), liver cancer, pancreatic cancer,ovarian cancer, kidney cancer; and decreasing or inhibiting tumorcellular proliferation or tumor growth in vivo. In one embodiment, ananti-CDCP1 antibody or ADC is capable of being internalized into a cellexpressing CDCP1 and/or inducing cytotoxicity.

The term “epitope” refers to a region of an antigen that is bound by anantibody, antibody fragment, or ADC. In certain embodiments, epitopedeterminants include chemically active surface groupings of moleculessuch as amino acids, sugar side chains, phosphoryl, or sulfonyl, and, incertain embodiments, may have specific three dimensional structuralcharacteristics, and/or specific charge characteristics. In certainembodiments, an antibody is said to specifically bind an antigen when itpreferentially recognizes its target antigen in a complex mixture ofproteins and/or macromolecules.

The term “surface plasmon resonance”, as used herein, refers to anoptical phenomenon that allows for the analysis of real-time biospecificinteractions by detection of alterations in protein concentrationswithin a biosensor matrix, for example using the BIAcore system(Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.). Forfurther descriptions, see Jönsson, U., et al. (1993) Ann. Biol. Clin.51:19-26; Jönsson, U., et al. (1991) Biotechniques 11:620-627; Johnsson,B., et al. (1995) J. Mol. Recognit. 8:125-131; and Johnnson, B., et al.(1991) Anal. Biochem. 198:268-277.

The term “k_(on)” or “k_(a)”, as used herein, is intended to refer tothe on rate constant for association of an antibody to the antigen toform the antibody/antigen complex.

The term “k_(off)” or “k_(d)”, as used herein, is intended to refer tothe off rate constant for dissociation of an antibody from theantibody/antigen complex.

The term “K_(D)”, as used herein, is intended to refer to theequilibrium dissociation constant of a particular antibody-antigeninteraction. K_(D) is calculated by k_(a)/k_(d). In one embodiment, theantibodies of the invention have a K_(D) of about 2,000 nM or less,about 1,000 nM or less, about 500 nM or less, about 200 nM or less,about 100 nM or less, about 75 nM or less, about 25 nM or less, about 21nM or less, about 12 nM or less, about 11 nM or less, about 10 nM orless, about 9 nM or less, about 8 nM or less, about 7 nM or less, about6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM orless, about 2 nM or less, about 1 nM or less, about 0.5 nM or less,about 0.3 nM or less, about 0.1 nM or less, about 0.01 nM or less, orabout 0.001 nM or less.

The term “competitive binding”, as used herein, refers to a situation inwhich a first antibody competes with a second antibody, for a bindingsite on a third molecule, e.g., an antigen. In one embodiment,competitive binding between two antibodies is determined using FACSanalysis.

The term “competitive binding assay” is an assay used to determinewhether two or more antibodies bind to the same epitope. In oneembodiment, a competitive binding assay is a competition fluorescentactivated cell sorting (FACS) assay which is used to determine whethertwo or more antibodies bind to the same epitope by determining whetherthe fluorescent signal of a labeled antibody is reduced due to theintroduction of a non-labeled antibody, where competition for the sameepitope will lower the level of fluorescence.

The term “labeled antibody” as used herein, refers to an antibody, or anantigen binding portion thereof, with a label incorporated that providesfor the identification of the binding protein, e.g., an antibody.Preferably, the label is a detectable marker, e.g., incorporation of aradiolabeled amino acid or attachment to a polypeptide of biotinylmoieties that can be detected by marked avidin (e.g., streptavidincontaining a fluorescent marker or enzymatic activity that can bedetected by optical or colorimetric methods). Examples of labels forpolypeptides include, but are not limited to, the following:radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In,¹²⁵I, ¹³¹I, ¹⁷⁷Lu, ¹⁶⁶Ho, or ¹⁵³Sm); fluorescent labels (e.g., FITC,rhodamine, lanthanide phosphors), enzymatic labels (e.g., horseradishperoxidase, luciferase, alkaline phosphatase); chemiluminescent markers;biotinyl groups; predetermined polypeptide epitopes recognized by asecondary reporter (e.g., leucine zipper pair sequences, binding sitesfor secondary antibodies, metal binding domains, epitope tags); andmagnetic agents, such as gadolinium chelates.

The term “antibody-drug-conjugate” or “ADC” refers to a binding protein,such as an antibody or antigen binding fragment thereof, chemicallylinked to one or more chemical drug(s) (also referred to herein asagent(s)) that may optionally be therapeutic or cytotoxic agents. In apreferred embodiment, an ADC includes an antibody, a cytotoxic ortherapeutic drug, and a linker that enables attachment or conjugation ofthe drug to the antibody. An ADC typically has anywhere from 1 to 8drugs conjugated to the antibody, including drug loaded species of 2, 4,6, or 8. Non-limiting examples of drugs that may be included in the ADCsare mitotic inhibitors, antitumor antibiotics, immunomodulating agents,vectors for gene therapy, alkylating agents, antiangiogenic agents,antimetabolites, boron-containing agents, chemoprotective agents,hormones, antihormone agents, corticosteroids, photoactive therapeuticagents, oligonucleotides, radionuclide agents, topoisomerase inhibitors,tyrosine kinase inhibitors, and radiosensitizers.

The term “antibody drug conjugate” refers to an ADC comprising anantibody, or antigen-binding portion thereof, that specifically binds toCDCP1, whereby the antibody is conjugated to one or more chemicalagent(s) or payloads. In one embodiment, the chemical agent is linked tothe antibody via a linker.

The term “drug-to-antibody ratio” or “DAR” refers to the number ofdrugs, e.g., IGN, auristatin, or maytansinoid, attached to the antibodyof the ADC. The DAR of an ADC can range from 1 to 8, although higherloads, e.g., 10, are also possible depending on the number of linkagesite on an antibody. The term DAR may be used in reference to the numberof drugs loaded onto an individual antibody, or, alternatively, may beused in reference to the average or mean DAR of a group of ADCs.

The term “CDCP1 associated disorder,” as used herein, includes anydisorder or disease (including proliferative disorders, e.g., cancer)that is marked, diagnosed, detected or identified by a phenotypic orgenotypic aberration of CDCP1 genetic components or expression duringthe course or etiology of the disease or disorder. In this regard aCDCP1 phenotypic aberration or determinant may, for example, compriseincreased or decreased levels of CDCP1 protein expression on one cellpopulation, e.g., a cancer cell population, as compared to another cellpopulation, e.g., a normal cell population, or increased or decreasedCDCP1 protein expression on certain definable cell populations, orincreased or decreased CDCP1 protein expression at an inappropriatephase or stage of a cell lifecycle. It will be appreciated that similarexpression patterns of genotypic determinants (e.g., mRNA transcriptionlevels) of CDCP1 may also be used to classify or detect CDCP1 associateddisorders. In one embodiment, an CDCP1 associated disorder is breastcancer, e.g., triple negative breast cancer. In another embodiment, anCDCP1 associated disorder is colon cancer. In another embodiment, aCDCP1 associated disorder is lung cancer, e.g., non-small cell lungcancer (NSCLC). In another embodiment, an CDCP1 associated disorder isliver cancer. In another embodiment, an CDCP1 associated disorder ispancreatic cancer. In another embodiment, an CDCP1 associated disorderis ovarian cancer. In another embodiment, an CDCP1 associated disorderis kidney cancer.

The term “cancer,” as used herein, is meant to refer to or describe thephysiological condition in mammals that is typically characterized byunregulated cell growth. Examples of cancer include, but are not limitedto, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoidmalignancies. More particular examples of such cancers include, but arenot limited to, breast cancer (Luminal A, TNBC, Ductal), prostatecancer, squamous cell tumors, squamous cell carcinoma (e.g., squamouscell lung cancer or squamous cell head and neck cancer), neuroendocrinetumors, urothelial cancer, vulvar cancer, mesothelioma, liver cancer,hone cancer, pancreatic cancer, skin cancer, cancer of the head or neck,lung cancer, small cell lung cancer, non-small cell lung cancer,cutaneous or intraocular malignant melanoma, renal cancer, uterinecancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer,cancer of the anal region, stomach cancer, testicular cancer, uterinecancer, carcinoma of the fallopian tubes, carcinoma of the endometrium,carcinoma of the cervix, carcinoma of the vagina, carcinoma of thevulva, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of thesmall intestine, cancer of the endocrine system, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, solid tumors of childhood,lymphocytic lymphoma, cancer of the bladder, cancer of the kidney orureter, carcinoma of the renal pelvis, neoplasm of the central nervoussystem (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axistumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma,epidermoid cancer, environmentally induced cancers including thoseinduced by asbestos, hematologic malignancies including, for example,multiple myeloma, B-cell lymphoma, Hodgkin lymphoma/primary mediastinalB-cell lymphoma, non-Hodgkin's lymphomas, acute myeloid leukemia (AML),myelodysplastic syndrome (MDS), acute lymphoblastic leukemia (ALL),chronic myelogenous leukemia (CML), myeloproliferative disorders (MPD),chronic lymphoid leukemia, follicular lymphoma, diffuse large B-celllymphoma, Burkitt's lymphoma, immunoblastic large cell lymphoma,precursor B-lymphoblastic lymphoma, mantle cell lymphoma, acutelymphoblastic leukemia, mycosis fungoides, anaplastic large celllymphoma, T-cell lymphoma, and precursor T-lymphoblastic lymphoma, PVNS,acute myeloid leukemia, adrenocortico carcinoma, ladder urothelialcarcinoma, cervical squamous cell carcinoma, endocervicaladenocarcinoma, diffuse large B cell lymphoma, glioblastoma multiforme,chronic lymphocytic leukemia, brain lower grade glioma, head and necksquamous cell carcinoma, hepatocellular carcinoma, lung adenocarcinoma,large squamous cell carcinoma, cutaneous melanoma, ovarial serouscystadenocarcinoma, gastric cancer, soft tissue sarcoma, testicular germcell cancer, thymoma, thyroid carcinoma, uterine corpus endometrialcarcinoma, uterine carcinosarcoma, kidney renal clear cell carcinoma,and kidney renal papillary cell carcinoma, and any combinations of saidcancers. The present invention is also applicable to treatment ofmetastatic cancers.

In one embodiment, the antibodies or ADCs of the invention areadministered to a patient having a solid tumor, including an advancedsolid tumor. In one embodiment, the antibodies or ADCs of the inventionare administered to a patient having a leukemia. In another embodiment,administration of antibodies or ADCs of the invention induce cell deathof CDCP1 expressing cells.

The term “CDCP1 expressing tumor,” as used herein, refers to a tumorwhich expresses CDCP1 protein (including a tumor comprising tumorinfiltrating cells that express CDCP1 protein). In one embodiment, CDCP1expression in a tumor is determined using immunohistochemical stainingof tumor cell membranes, where any immunohistochemical staining abovebackground level in a tumor sample indicates that the tumor is a CDCP1expressing tumor. In another embodiment, a CDCP1 expressing tumor isidentified in a patient when greater than 1%, greater than 2%, greaterthan 3%, greater than 4%, greater than 5%, greater than 6%, greater than7%, greater than 8%, greater than 9%, greater than 10%, greater than15%, greater than 20%, greater than 25%, or greater than 30%, greaterthan 40%, greater than 50%, greater than 60%, greater than 70%, greaterthan 80%, greater than 90%, or more of the cells in a tumor sample arepositive for CDCP1 expression. In another embodiment, CDCP1 positiveexpression is determined based on membrane staining as determined by,e.g., immunohistochemistry (IHC) analysis.

A CDCP1 expressing tumor is identified as having an “elevated level ofCDCP1” or “expressing CDCP1 at an elevated level” when the level ofCDCP1 is higher than in tissue surrounding the cancer. In someembodiments, an “elevated level of CDCP1” is one in which 5% or more ofthe cells in a tumor sample have membrane staining. In some embodimentsa “high level” in regard to CDCP1 is 5%) or more staining, for example,5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100% of the cells in the tumorsample are stained. In some embodiments, the protein expression levelscan be measured by IHC analysis.

A CDCP1 expressing tumor is identified as having a “low level of CDCP1”or “expressing CDCP1 at a low level” is one in which 5% or less of thecells in a tumor sample have membrane staining. In some embodiments a“low level” in regard to CDCP1 is 5% or less staining, for example, 4.9,4.5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1% or less ofthe cells in the tumor sample are stained. In some embodiments, theprotein expression levels can be measured by IHC analysis.

A cell that expresses no CDCP1 can also be described as expressing a“low level of CDCP1”. Thus, the phrase “expresses a low level of CDCP1”encompasses no CDCP1 expression. In some embodiments, a low level ofCDCP1 is within the background staining levels. In some embodiments, asample that is CDCP1 “negative” has no CDCP1 expression or a low levelof CDCP1. In some embodiments, CDCP1 staining is negative when no orless than 5%, 4%, 3%, 2%, or 1% of the cells have membrane staining forCDCP1.

As used herein, the term “tumor sample” refers to a tumor tissue or cellsample obtained from a solid tumor. The sample can include both tumorcells and tumor infiltrating cells, e.g., tumor infiltrating immunecells.

As used herein, the term “non-cancer sample” or “normal sample” refersto a sample from a normal tissue (e.g., a lung or ovarian tissue sampleor a normal cell sample). In some embodiments, the non-cancer samplecomes from the same subject, but is from a different part of the subjectthan that being tested. In some embodiments, the non-cancer sample isfrom a tissue area surrounding or adjacent to the cancer. In someembodiments, the non-cancer sample is not from the subject being tested,but is a sample from a subject known to have, or not to have, a disorderin question (for example, a particular cancer such a CDCP1 relateddisorder). In some embodiments, the non-cancer sample is from the samesubject, but from a point in time before the subject developed cancer.In some embodiments, the reference sample is from a benign cancer sample(for example, benign ovarian cancer sample), from the same or adifferent subject.

Methods for detecting expression of CDCP1 in a tumor are known in theart.

The terms “overexpress,” “overexpression,” or “overexpressed”interchangeably refer to a gene that is transcribed or translated at adetectably greater level, usually in a cancer cell, in comparison to anormal cell. Overexpression therefore refers to both overexpression ofprotein and RNA (due to increased transcription, post transcriptionalprocessing, translation, post translational processing, alteredstability, and altered protein degradation), as well as localoverexpression due to altered protein traffic patterns (increasednuclear localization), and augmented functional activity, e.g., as in anincreased enzyme hydrolysis of substrate. Thus, overexpression refers toeither protein or RNA levels. Overexpression can also be by 5%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in comparison to a normalcell or comparison cell. In certain embodiments, the anti-CDCP1antibodies or ADCs are used to treat solid tumors likely to overexpressCDCP1.

The term “administering” as used herein is meant to refer to thedelivery of a substance (e.g., an anti-CDCP1 antibody or ADC) to achievea therapeutic objective (e.g., the treatment of an CDCP1-associateddisorder or the inhibition or reduction of a tumor). Modes ofadministration may be parenteral, enteral and topical. Parenteraladministration is usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraartcrial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The term “combination therapy”, as used herein, refers to theadministration of two or more therapeutic substances, e.g., ananti-CDCP1 antibody or ADC and an additional therapeutic agent. Theadditional therapeutic agent may be administered concomitant with, priorto, or following the administration of the anti-CDCP1 antibody or ADC.In one embodiment, the anti-CDCP1 antibodies or ADCs of the inventionare administered in combination with one or more immune checkpointinhibitors or (e.g., one or more antibody or small molecule immunecheckpoint inhibitors) for the treatment of a cancer. In one embodiment,the anti-CDCP1 antibodies, bispecific antibodies, or ADCs disclosedherein are administered in combination with a PARP (poly ADP ribosepolymerase) inhibitor. PARP inhibitors are well known to those ofordinary skill in the art and include, but are not limited to,Niraparib, Olaparib, Rucaparib, Iniparib, Talazoparib, Vcliparib, CEP9722, E7016, BGB-290, and 3-aminobenazamine.

As used herein, the term “effective amount” or “therapeuticallyeffective amount” refers to the amount of a drug, e.g., an antibody orADC, which is sufficient to reduce or ameliorate the severity and/orduration of a disorder, e.g., cancer, or one or more symptoms thereof,prevent the advancement of a disorder, cause regression of a disorder,prevent the recurrence, development, onset or progression of one or moresymptoms associated with a disorder, detect a disorder, or enhance orimprove the prophylactic or therapeutic effect(s) of another therapy(e.g., prophylactic or therapeutic agent). The effective amount of anantibody or ADC may, for example, inhibit tumor growth (e.g., inhibit anincrease in tumor volume), decrease tumor growth (e.g., decrease tumorvolume), reduce the number of cancer cells, and/or relieve to someextent one or more of the symptoms associated with the cancer. Theeffective amount may, for example, improve disease free survival (DFS),improve overall survival (OS), or decrease likelihood of recurrence.

Various aspects of the invention are described in further detail in thefollowing subsections.

II. Anti-CDCP1 Antibodies

One aspect disclosed herein provides humanized anti-CDCP1 antibodies, orantigen binding portions thereof. Another aspect disclosed hereinprovides human anti-CDCP1 antibodies, or antigen binding portionsthereof. In one embodiment, the antibodies disclosed herein bind humanCDCP1. In another embodiment, the antibodies disclosed herein bindcynomolgus monkey CDCP1. In another embodiment, the antibodies disclosedherein bind human CDCP1 expressed on tumor cells.

Another aspect disclosed herein features antibody drug conjugates (ADCs)comprising an anti-CDCP1 antibody described herein and at least onedrug(s). The antibodies or ADCs disclosed herein have characteristicsincluding, but not limited to, binding to human CDCP1 in vitro,modulating, e.g., inhibiting IL-1 signaling, inducing cell death incells expressing CDCP1, including, but not limited to, leukemia cells,and decreasing or inhibiting cancer, tumor cellular proliferation ortumor growth, or tumor invasion and metastasis. ADCs disclosed herein,in particular, have characteristics including, but not limited to,inducing cell death in cells expressing CDCP1, e.g., leukemia cellsexpressing CDCP1. In one embodiment, an anti-CDCP1 antibody or ADCdisclosed herein is capable of being internalized into a cell expressingCDCP1.

In one embodiment, anti-CDCP1 antibodies are disclosed which have theability to bind to CDCP1, as described in the Examples below.Collectively, the novel antibodies are referred to herein as “CDCP1antibodies.” The anti-CDCP1 antibodies, ADCs, or antigen bindingfragments thereof, are able to inhibit or decrease tumor growth in vivo.The tumor can be a CDCP1 negative tumor or an CDCP1 expressing tumor. Invarious embodiments, anti-CDCP1 antibodies, ADCs, or antigen bindingfragments thereof, are capable of modulating a biological function ofCDCP1. In other embodiments of the foregoing aspects, the anti-CDCP1antibodies, ADCs, or antigen binding fragments thereof, bind CDCP1 oncells expressing CDCP1. Thus, the disclosure includes anti-CDCP1antibodies, ADCs, or antigen binding fragments thereof, that areeffective at inhibiting or decreasing tumor growth.

In addition, the present inventors have shown that CDCP1 is expressed bynumerous solid tumors, including breast cancer tumors, e.g., triplenegative breast cancer tumors, lung cancer tumors, e.g., non-small celllung cancer tumors, liver cancer tumors, pancreatic cancer tumors,ovarian cancer tumors, kidney cancer tumors, and colon cancer tumors(see, e.g., Examples 3-7). Accordingly, the anti-CDCP1 antibodies, ADCs,and antigen-binding portions thereof, can be used for the treatment ofbreast cancer, e.g., triple negative breast cancer, lung cancer, e.g.,non-small cell lung cancer, liver cancer, pancreatic cancer, ovariancancer, kidney cancer, and colon cancer in a subject. In one embodiment,greater than 1%, greater than 2%, greater than 3%, greater than 4%,greater than 5%, greater than 6%, greater than 7%, greater than 8%,greater than 9%, greater than 10%, greater than 15%, greater than 20%,greater than 25%, or greater than 30%, greater than 40%, greater than50%, greater than 60%, greater than 70%, greater than 80%, greater than90%, or more of the cells in a tumor sample, e.g., breast cancer tumor,e.g., triple negative breast cancer tumor, lung cancer tumor, e.g.,non-small cell lung cancer tumor, liver cancer tumor, pancreatic cancertumor, ovarian cancer tumor, kidney cancer tumor, and colon cancertumor, are positive for CDCP1 expression. In another embodiment, a tumorsample has a high level of CDCP1 expression. For example, in oneembodiment, at least 5% or more of the cells in a tumor sample, e.g.,breast cancer tumor, e.g., triple negative breast cancer tumor, lungcancer tumor, e.g., non-small cell lung cancer tumor, liver cancertumor, pancreatic cancer tumor, ovarian cancer tumor, kidney cancertumor, and colon cancer tumor, have membrane staining. In anotherembodiment, a tumor sample obtained from the subject displays a lowlevel of expression of CDCP1. The expression level of CDCP1 can bedetermined by any method known in the art. For example, the expressionlevel of CDCP1 can be determined via immunohistochemical analysis. Inanother embodiment, the cancer has been previously treated with anotheranti-cancer agent or anti-cancer therapy, e.g., a chemotherapy. In oneembodiment, the cancer is resistant to chemotherapy.

Antibodies having combinations of any of the aforementionedcharacteristics are contemplated as aspects of the disclosure. ADCs,described in more detail below, may also have any of the foregoingcharacteristics.

One aspect of the disclosure features an anti-human CDCP1 (anti-hCDCP1)Antibody Drug Conjugate (ADC) comprising an anti-hCDCP1 antibodyconjugated to a drug via a linker. Exemplary anti-CDCP1 antibodies (andsequences thereof) that can be used in the ADCs are described herein.

The anti-CDCP1 antibodies described herein provide the ADCs with theability to bind to CDCP1 such that the cytotoxic molecule attached tothe antibody may be delivered to the CDCP1-expressing cell, particularlya CDCP1 expressing cancer cell.

While the term “antibody” is used throughout, it should be noted thatantibody fragments (i.e., antigen-binding portions of an anti-CDCP1antibody) are also included in the disclosure and may be included in theembodiments (methods and compositions) described throughout. Forexample, an anti-CDCP1 antibody fragment may be conjugated to the drugs,as described herein. In certain embodiments, an anti-CDCP1 antibodybinding portion is a Fab, a Fab′, a F(ab′)2, a Fv, a disulfide linkedFv, an scFv, a single domain antibody, or a diabody.

Example 2 describes the generation of fully human CDCP1 antibodiesagainst the extracellular domain of human CDCP1. The heavy and lightchain variable region CDR amino acid sequences and the heavy and lightchain variable region amino acid sequences for these human antibodiesare set forth in FIGS. 15, 18, and 20 , and the the nucleotide sequenceencoding the heavy and light chain variable region amino acid sequencesfor these human antibodies are set forth in FIG. 19 .

Thus, in one embodiment, the disclosure includes human anti-CDCP1antibodies, or antigen binding portions thereof, comprising a heavychain variable region comprising an amino acid sequence selected fromthe group consisting of SEQ ID NOs:1, 11, 21, 31, 41, 51, 61, 71, 81,91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221,231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361,371, 381, and 391; and a light chain variable region comprising an aminoacid sequence selected from the group consisting of 2, 12, 22, 32, 42,52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192,202, 222, 222, 232, 242, 252, 262, 272, 282, 292, 302, 322, 322, 332,342, 352, 362, 372, 382, and 392.

In one embodiment, the disclosure includes a human anti-CDCP1 antibody,or antigen binding portion thereof, comprising an FIC CDR set (CDR1,CDR2, and CDR3) selected from those set forth in FIG. 15 or 20 ; and anLC CDR set (CDR1, CDR2, and CDR3) selected from those set forth in FIG.15 or 20 .

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 38E11. The 38E11 antibody comprises aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 157, a CDR2 domain comprising theamino acid sequence of SEQ ID NO: 156, and a CDR1 domain comprising theamino acid sequence of SEQ ID NO: 155, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 160, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:159, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:158. In further embodiments, disclosed herein is an antibody having aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 151 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 152.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 151, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 151, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 152, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 152.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 27H10. The 27H10 antibody comprises aheavy chain variable region comprising a CDR3 domain comprising theamino acid sequence of SEQ ID NO: 37, a CDR2 domain comprising the aminoacid sequence of SEQ ID NO: 36, and a CDR1 domain comprising the aminoacid sequence of SEQ ID NO: 35, and a light chain variable regioncomprising a CDR3 domain comprising the amino acid sequence of SEQ IDNO: 40, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:39, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:38. In further embodiments, disclosed herein is an antibody having aheavy chain variable region comprising the amino acid sequence of SEQ IDNO: 31 and a light chain variable region comprising the amino acidsequence of SEQ ID NO: 32.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 31, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 31, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 32, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 32.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 18C6. The 18C6 antibody comprises a heavychain variable region comprising a CDR3 domain comprising the amino acidsequence of SEQ ID NO: 87, a CDR2 domain comprising the amino acidsequence of SEQ ID NO: 86, and a CDR1 domain comprising the amino acidsequence of SEQ ID NO: 85, and a light chain variable region comprisinga CDR3 domain comprising the amino acid sequence of SEQ ID NO: 90, aCDR2 domain comprising the amino acid sequence of SEQ ID NO: 89, and aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 88. Infurther embodiments, disclosed herein is an antibody having a heavychain variable region comprising the amino acid sequence of SEQ ID NO:81 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 82.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 81, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 81, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 82, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 82.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 10E2. The 10E2 antibody comprises a heavychain variable region comprising a CDR3 domain comprising the amino acidsequence of SEQ ID NO: 107, a CDR2 domain comprising the amino acidsequence of SEQ ID NO: 106, and a CDR1 domain comprising the amino acidsequence of SEQ ID NO: 105, and a light chain variable region comprisinga CDR3 domain comprising the amino acid sequence of SEQ ID NO: 110, aCDR2 domain comprising the amino acid sequence of SEQ ID NO: 109, and aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 108. Infurther embodiments, disclosed herein is an antibody having a heavychain variable region comprising the amino acid sequence of SEQ ID NO:101 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 102.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 101, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 101, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 102, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 102.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 41A9. The 41A9 antibody comprises a heavychain variable region comprising a CDR3 domain comprising the amino acidsequence of SEQ ID NO: 127, a CDR2 domain comprising the amino acidsequence of SEQ ID NO: 126, and a CDR1 domain comprising the amino acidsequence of SEQ ID NO: 125, and a light chain variable region comprisinga CDR3 domain comprising the amino acid sequence of SEQ ID NO: 130, aCDR2 domain comprising the amino acid sequence of SEQ ID NO: 129, and aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 128. Infurther embodiments, disclosed herein is an antibody having a heavychain variable region comprising the amino acid sequence of SEQ ID NO:121 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 122.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 121, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 121, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 122, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 122.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 3B11. The 3B11 antibody comprises a heavychain variable region comprising a CDR3 domain comprising the amino acidsequence of SEQ ID NO: 47, a CDR2 domain comprising the amino acidsequence of SEQ ID NO: 46, and a CDR1 domain comprising the amino acidsequence of SEQ ID NO: 45, and a light chain variable region comprisinga CDR3 domain comprising the amino acid sequence of SEQ ID NO: 50, aCDR2 domain comprising the amino acid sequence of SEQ ID NO: 49, and aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 48. Infurther embodiments, disclosed herein is an antibody having a heavychain variable region comprising the amino acid sequence of SEQ ID NO:41 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 42.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 41, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 41, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 42, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 42.

In one embodiment, an anti-CDCP1 antibody, or antigen binding portionthereof, is the human antibody 47G7. The 47G7 antibody comprises a heavychain variable region comprising a CDR3 domain comprising the amino acidsequence of SEQ ID NO: 77, a CDR2 domain comprising the amino acidsequence of SEQ ID NO: 76, and a CDR1 domain comprising the amino acidsequence of SEQ ID NO: 75, and a light chain variable region comprisinga CDR3 domain comprising the amino acid sequence of SEQ ID NO: 80, aCDR2 domain comprising the amino acid sequence of SEQ ID NO: 79, and aCDR1 domain comprising the amino acid sequence of SEQ ID NO: 78. Infurther embodiments, disclosed herein is an antibody having a heavychain variable region comprising the amino acid sequence of SEQ ID NO:71 and a light chain variable region comprising the amino acid sequenceof SEQ ID NO: 72.

In some embodiments, an anti-CDCP1 antibody, or antigen-binding portionthereof, comprises a heavy chain comprising an amino acid sequence setforth in SEQ ID NO: 71, or a sequence having at least 90%, 95%, 96%,97%, 98%, or 99% identity to SEQ ID NO: 71, and/or a light chaincomprising an amino acid sequence set forth in SEQ ID NO: 72, or asequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQID NO: 72.

The foregoing anti-CDCP1 antibody CDR sequences establish a novel familyof CDCP1 binding proteins, isolated in accordance with this disclosure,and comprising antigen binding polypeptides that include the CDRsequences listed in FIGS. 15, 18, and 20 , as well as the Sequencesummary.

In some embodiments, the anti-CDCP1 antibodies of the present inventionare human or humanized. In some embodiments, the anti-CDCP1 antibodiesspecifically bind to the same epitope as the specific antibodiesdisclosed herein.

The disclosure also provides antibodies that specifically bind tospecific epitopes on a target CDCP1 molecule. Likewise, the disclosureprovides epitopes useful for identifying the antibodies thatspecifically bind to a target CDCP1 molecule comprising the epitope.

Epitope mapping can be done using standard methods. For example, phagedisplay is an in vitro selection technique in which a peptide isgenetically fused to a coat protein of a bacteriophage resulting indisplay of a fused protein on the exterior of the virion. Biopanning ofthese virions by incubating the pool of phage displayed variants with aspecific antibody of interest, which has been immobilized on a plate.The unbound phage is then washed away and the specifically bound phageis then eluted. The eluted phage is then amplified in E. coli and theprocess is repeated, resulting in enrichment of the phage pool in favorof the tightest binding sequences.

An advantage of this technology is that it allows for the screening ofgreater than 10⁹ sequences in an unbiased way. Phage display isespecially useful if the immunogen is unknown or a large proteinfragment. One of the limitations of phage display includescross-contamination between phage particles that can enrich sequencesthat do not specifically bind to the antibody. Additionally, sequencesthat are not found in nature will be present in the phage displayedpeptide library. These sequences may not resemble the immunizing peptideat all and may bind tightly to the antibody of interest. Retrievingsequences that do not resemble the immunizing peptide can be veryconfounding and it is difficult to decipher whether these peptides arecontamination or unnatural peptides with high binding affinity to theantibody of interest.

In another aspect, the present disclosure features polynucleotidesequences that comprise or consist of the sequences as shown in FIG. 19, and/or amino acid sequences that comprise or consist of the sequencesas shown in FIG. 18 and FIG. 20 .

In one aspect, the disclosure relates to an antibody or antigen-bindingfragment thereof that binds to CUB domain-containing protein 1 (CDCP1)having a heavy chain variable region (VH) including complementaritydetermining regions (CDRs) 1, 2, 3, wherein the CDR1 region includes anamino acid sequence that is at least 80%, 90%, 92%, 95%, 97%, 98%, 99%,or 100% identical to a selected VH CDR1 amino acid sequence, the CDR2region includes an amino acid sequence that is at least 80%, 90%, 92%,95%, 97%, 98%, 99%, or 100% identical to a selected VH CDR2 amino acidsequence, and the CDR3 region includes an amino acid sequence that is atleast 80%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to a selectedVH CDR3 amino acid sequence, and a light chain variable region (VL)comprising CDRs 1, 2, 3, wherein the CDR1 region comprises an amino acidsequence that is at least 80%, 90%, 92%, 95%, 97%, 98%, 99%, or 100%identical to a selected VL CDR1 amino acid sequence, the CDR2 regioncomprises an amino acid sequence that is at least 80%, 90%, 92%, 95%,97%, 98%, 99%, or 100% identical to a selected VL CDR2 amino acidsequence, and the CDR3 region comprises an amino acid sequence that isat least 80%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to aselected VL CDR3 amino acid sequence, wherein the selected VH CDRs 1, 2,3 amino acid sequences and the selected VL CDRs, 1, 2, 3 amino acidsequences are one of the following:

(1) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 5, 6, 7, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 8, 9, 10,respectively;

(2) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 15, 16, 17, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 18, 19, 20,respectively;

(3) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 25, 26, 27, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 28, 29, 30,respectively;

(4) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 35, 36, 37, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 38, 39, 40,respectively;

(5) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 45, 46, 47, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 48, 49, 50,respectively;

(6) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 55, 56, 57, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 58, 59, 60,respectively;

(7) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 65, 66, 67, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 68, 69, 70,respectively;

(8) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 75, 76, 77, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 78, 79, 80,respectively;

(9) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 85, 86, 87, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 88, 89, 90,respectively;

(10) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 95, 96, 97, respectively, and the selected VL CDRs 1, 2, 3amino acid sequences are set forth in SEQ ID NOs: 98, 99, 100,respectively;

(11) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 105, 106, 107, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 108, 109, 110,respectively;

(12) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 115, 116, 117, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 118, 119, 120,respectively;

(13) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 125, 126, 127, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 128, 129, 130,respectively;

(14) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 135, 136, 137, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 138, 139, 140,respectively;

(15) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 145, 146, 147, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 148, 149, 150,respectively;

(16) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 155, 156, 157, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 158, 159, 160,respectively;

(17) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 165, 166, 167, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 168, 169, 170,respectively;

(18) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 175, 176, 177, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 178, 179, 180,respectively;

(19) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 185, 186, 187, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 188, 189, 190,respectively;

(20) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 195, 196, 197, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 198, 199, 200,respectively;

(21) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 205, 206, 207, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 208, 209, 210,respectively;

(22) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 215, 216, 217, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 218, 219, 220,respectively;

(23) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 225, 226, 227, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 228, 229, 230,respectively;

(24) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 235, 236, 237, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 238, 239, 240,respectively:

(25) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 245, 246, 247, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 248, 249, 250,respectively;

(26) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 255, 256, 257, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 258, 259, 260,respectively;

(27) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 265, 266, 267, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 268, 269, 270,respectively:

(28) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 275, 276, 277, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 278, 279, 280,respectively;

(29) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 285, 286, 287, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 288, 289, 290,respectively;

(30) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 295, 296, 297, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 298, 299, 300,respectively;

(31) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 305, 306, 307, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 308, 309, 310,respectively;

(32) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 315, 316, 317, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 318, 319, 320,respectively;

(33) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 325, 326, 327, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 328, 329, 330,respectively;

(34) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 335, 336, 337, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 338, 339, 340,respectively;

(35) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 345, 346, 347, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 348, 349, 350,respectively;

(36) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 355, 356, 357, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 358, 359, 360,respectively;

(37) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 365, 366, 367, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 368, 369, 370,respectively;

(38) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 375, 376, 377, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 378, 379, 380,respectively;

(39) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 385, 386, 387, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 388, 389, 390,respectively; and

(40) the selected VH CDRs 1, 2, 3 amino acid sequences are set forth inSEQ ID NOs: 395, 396, 397, respectively, and the selected VL CDRs 1, 2,3 amino acid sequences are set forth in SEQ ID NOs: 398, 399, 400,respectively,

wherein the antibody or antigen-binding fragment thereof specificallybinds to CUB domain-containing protein 1 (CDCP1).

In some embodiments, the antibody or antigen-binding fragmentspecifically binds to human CDCP1 and/or Cynomolgus CDCP1.

In some embodiments, the antibody or antigen-binding fragment has adissociation constant (K_(d)) for human CDCP1 that is less than 10 nM,and/or a K_(d) for Cynomolgus CDCP1 that is less than 10 nM.

In some embodiments, the antibody or antigen-binding fragment is ahumanized antibody or antigen-binding fragment thereof.

The disclosure also provides a cDNA including a polynucleotide encodinga polypeptide comprising:

(1) an immunoglobulin heavy chain or a fragment thereof comprising aheavy chain variable region (VH) comprising complementarity determiningregions (CDRs) 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 5, 6, and 7, respectively, and wherein the VH when pairedwith a light chain variable region (VL) comprising the amino acidsequence set forth in SEQ ID NO: 2 binds to CDCP1;

(2) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 8, 9, and 10, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO:1binds to CDCP1;

(3) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 15, 16, and 17, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 12binds to CDCP1;

(4) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 18, 19, and 20, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 11binds to CDCP1;

(5) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 25, 26, and 27, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 22binds to CDCP1;

(6) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 28, 29, and 30, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 21binds to CDCP1;

(7) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 35, 36, and 37, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 32binds to CDCP1;

(8) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 38, 39, and 40, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 31binds to CDCP1;

(9) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 45, 46, and 47, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 42binds to CDCP1;

(10) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 48, 49, and 50, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 41binds to CDCP1;

(11) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 55, 56, and 57, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 52binds to CDCP1;

(12) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 58, 59, and 60, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 51binds to CDCP1;

(13) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 65, 66, and 67, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 62binds to CDCP1;

(14) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 68, 69, and 70, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 61binds to CDCP1;

(15) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 75, 76, and 77, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 72binds to CDCP1;

(16) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 78, 79, and 80, respectively, and wherein the VL when pairedwith a Vii comprising the amino acid sequence set forth in SEQ ID NO: 71binds to CDCP1;

(17) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 85, 86, and 87, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 82binds to CDCP1;

(18) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 88, 89, and 90, respectively, and wherein the VL when pairedwith a VH comprising the amino acid sequence set forth in SEQ ID NO: 81binds to CDCP1;

(19) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 95, 96, and 97, respectively, and wherein the VH when pairedwith a VL comprising the amino acid sequence set forth in SEQ ID NO: 92binds to CDCP1;

(20) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 98, 99, and 100, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 91 binds to CDCP1;

(21) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 105, 106, and 107, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 102 binds to CDCP1;

(22) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 108, 109, and 110, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 101 binds to CDCP1;

(23) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 115, 116, and 117, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 112 binds to CDCP1;

(24) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 118, 119, and 120, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 111 binds to CDCP1;

(25) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 125, 126, and 127, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 122 binds to CDCP1;

(26) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 128, 129, and 130, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 121 binds to CDCP1;

(27) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 135, 136, and 137, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 132 binds to CDCP1;

(28) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 138, 139, and 140, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 131 binds to CDCP1;

(29) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 145, 146, and 147, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 142 binds to CDCP1;

(30) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 148, 149, and 150, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 141 binds to CDCP1;

(31) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 155, 156, and 157, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 152 binds to CDCP1;

(32) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 158, 159, and 160, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 151 binds to CDCP1;

(33) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 165, 166, and 167, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 162 binds to CDCP1;

(34) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 168, 169, and 170, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 161 binds to CDCP1;

(35) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 175, 176, and 177, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 172 binds to CDCP1;

(36) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 178, 179, and 180, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 171 binds to CDCP1;

(37) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 185, 186, and 187, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 182 binds to CDCP1;

(38) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 188, 189, and 190, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 181 binds to CDCP1;

(39) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 195, 196, and 197, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 192 binds to CDCP1;

(40) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 198, 199, and 200, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 191 binds to CDCP1;

(41) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 205, 206, and 207, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 202 binds to CDCP1;

(42) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 208, 209, and 210, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 201 binds to CDCP1;

(43) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 215, 216, and 217, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 212 binds to CDCP1;

(44) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 218, 219, and 220, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 211 binds to CDCP1;

(45) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 225, 226, and 227, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 222 binds to CDCP1;

(46) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 228, 229, and 230, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 221 binds to CDCP1;

(47) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 235, 236, and 237, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 232 binds to CDCP1;

(48) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 238, 239, and 240, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 231 binds to CDCP1;

(49) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 245, 246, and 247, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 242 binds to CDCP1;

(50) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 248, 249, and 250, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 241 binds to CDCP1;

(51) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 255, 256, and 257, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 252 binds to CDCP1;

(52) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 258, 259, and 260, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 251 binds to CDCP1;

(53) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 265, 266, and 267, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 262 binds to CDCP1;

(54) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 268, 269, and 270, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 261 binds to CDCP1;

(55) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 275, 276, and 277, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 272 binds to CDCP1;

(56) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 278, 279, and 280, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 271 binds to CDCP1;

(57) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 285, 286, and 287, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 282 binds to CDCP1;

(58) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 288, 289, and 290, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 281 binds to CDCP1;

(59) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 295, 296, and 297, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 292 binds to CDCP1;

(60) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 298, 299, and 300, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 291 binds to CDCP1;

(61) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 305, 306, and 307, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 302 binds to CDCP1;

(62) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 308, 309, and 310, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 301 binds to CDCP1;

(63) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 315, 316, and 317, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 312 binds to CDCP1;

(64) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 318, 319, and 320, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 311 binds to CDCP1;

(65) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 325, 326, and 327, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 322 binds to CDCP1;

(66) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 328, 329, and 330, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 321 binds to CDCP1;

(67) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 335, 336, and 337, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 332 binds to CDCP1;

(68) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 338, 339, and 340, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 331 binds to CDCP1;

(69) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 345, 346, and 347, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 342 binds to CDCP1;

(70) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 348, 349, and 350, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 341 binds to CDCP1;

(71) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 355, 356, and 357, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 352 binds to CDCP1;

(72) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 358, 359, and 360, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 351 binds to CDCP1;

(73) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 365, 366, and 367, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 362 binds to CDCP1;

(74) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 368, 369, and 370, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 361 binds to CDCP1;

(75) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 375, 376, and 377, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 372 binds to CDCP1;

(76) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 378, 379, and 380, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 371 binds to CDCP1;

(77) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 385, 386, and 387, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 382 binds to CDCP1;

(78) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 388, 389, and 390, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 381 binds to CDCP1;

(79) an immunoglobulin heavy chain or a fragment thereof comprising a VHcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 395, 396, and 397, respectively, and wherein the VH whenpaired with a VL comprising the amino acid sequence set forth in SEQ IDNO: 392 binds to CDCP1; or

(80) an immunoglobulin light chain or a fragment thereof comprising a VLcomprising CDRs 1, 2, and 3 with the amino acid sequences set forth inSEQ ID NOs: 398, 399, and 400, respectively, and wherein the VL whenpaired with a VH comprising the amino acid sequence set forth in SEQ IDNO: 391 binds to CDCP1.

In some embodiments, the VH when paired with a VL specifically binds tohuman CDCP1 and/or Cynomolgus CDCP1, and the VL when paired with a VHspecifically binds to human CDCP1 and/or Cynomolgus CDCP1.

In some embodiments, the immunoglobulin heavy chain or the fragmentthereof is a humanized immunoglobulin heavy chain or a fragment thereof,and the immunoglobulin light chain or the fragment thereof is ahumanized immunoglobulin light chain or a fragment thereof.

In another aspect, the disclosure provides an antibody orantigen-binding fragment thereof that binds to CDCP1 having a heavychain variable region (VH) comprising an amino acid sequence that is atleast 80%, 90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to a selectedVH sequence, and a light chain variable region (VL) comprising an aminoacid sequence that is at least 80%, 90%, 92%, 95%, 97%, 98%, 99%, or100% identical to a selected VL sequence, wherein the selected VHsequence and the selected VL sequence are one of the following:

(1) the selected VH sequence is SEQ ID NO: 1, and the selected VLsequence is SEQ ID NO: 2;

(2) the selected VH sequence is SEQ ID NO: 11, and the selected VLsequence is SEQ ID NO: 12;

(3) the selected VH sequence is SEQ ID NO: 21, and the selected VLsequence is SEQ ID NO: 22;

(4) the selected VH sequence is SEQ ID NO: 31, and the selected VLsequence is SEQ ID NO: 32;

(5) the selected VH sequence is SEQ ID NO: 41, and the selected VLsequence is SEQ ID NO: 42;

(6) the selected VH sequence is SEQ ID NO: 51, and the selected VLsequence is SEQ ID NO: 52;

(7) the selected VH sequence is SEQ ID NO: 61, and the selected VLsequence is SEQ ID NO: 62;

(8) the selected VH sequence is SEQ ID NO: 71, and the selected VLsequence is SEQ ID NO: 72;

(9) the selected VH sequence is SEQ ID NO: 81, and the selected VLsequence is SEQ ID NO: 82;

(10) the selected VH sequence is SEQ ID NO: 91, and the selected VLsequence is SEQ ID NO: 92;

(11) the selected VH sequence is SEQ ID NO: 101, and the selected VLsequence is SEQ ID NO: 102;

(12) the selected VH sequence is SEQ ID NO: 111, and the selected VLsequence is SEQ ID NO: 112;

(13) the selected VH sequence is SEQ ID NO: 121, and the selected VLsequence is SEQ ID NO: 122;

(14) the selected VH sequence is SEQ ID NO: 131, and the selected VLsequence is SEQ ID NO: 132;

(15) the selected VH sequence is SEQ ID NO: 141, and the selected VLsequence is SEQ ID NO: 142;

(16) the selected VH sequence is SEQ ID NO: 151, and the selected VLsequence is SEQ ID NO: 152;

(17) the selected VH sequence is SEQ ID NO: 161, and the selected VLsequence is SEQ ID NO: 162; and

(18) the selected VH sequence is SEQ ID NO: 171, and the selected VLsequence is SEQ ID NO: 172;

(19) the selected VH sequence is SEQ ID NO: 181, and the selected VLsequence is SEQ ID NO: 182;

(20) the selected VH sequence is SEQ ID NO: 191, and the selected VLsequence is SEQ ID NO: 192;

(21) the selected VH sequence is SEQ ID NO: 201, and the selected VLsequence is SEQ ID NO: 202;

(22) the selected VH sequence is SEQ ID NO: 211, and the selected VLsequence is SEQ ID NO: 212;

(23) the selected VH sequence is SEQ ID NO: 221, and the selected VLsequence is SEQ ID NO: 222;

(24) the selected VH sequence is SEQ ID NO: 231, and the selected VLsequence is SEQ ID NO: 232;

(25) the selected VH sequence is SEQ ID NO: 241, and the selected VLsequence is SEQ ID NO: 242;

(26) the selected VH sequence is SEQ ID NO: 251, and the selected VLsequence is SEQ ID NO: 252;

(27) the selected VH sequence is SEQ ID NO: 261, and the selected VLsequence is SEQ ID NO: 262;

(28) the selected VH sequence is SEQ ID NO: 271, and the selected VLsequence is SEQ ID NO: 272;

(29) the selected VH sequence is SEQ ID NO: 281, and the selected VLsequence is SEQ ID NO: 282;

(30) the selected VH sequence is SEQ ID NO: 291, and the selected VLsequence is SEQ ID NO: 292;

(31) the selected VH sequence is SEQ ID NO: 301, and the selected VLsequence is SEQ ID NO: 302;

(32) the selected VH sequence is SEQ ID NO: 311, and the selected VLsequence is SEQ ID NO: 312;

(33) the selected VH sequence is SEQ ID NO: 321, and the selected VLsequence is SEQ ID NO: 322;

(34) the selected VH sequence is SEQ ID NO: 331, and the selected VLsequence is SEQ ID NO: 332;

(35) the selected VH sequence is SEQ ID NO: 341, and the selected VLsequence is SEQ ID NO: 342;

(36) the selected VH sequence is SEQ ID NO: 351, and the selected VLsequence is SEQ ID NO: 352;

(37) the selected VH sequence is SEQ ID NO: 361, and the selected VLsequence is SEQ ID NO: 362;

(38) the selected VH sequence is SEQ ID NO: 371, and the selected VLsequence is SEQ ID NO: 372;

(39) the selected VH sequence is SEQ ID NO: 381, and the selected VLsequence is SEQ ID NO: 382; and

(40) the selected VH sequence is SEQ ID NO: 391, and the selected VLsequence is SEQ ID NO: 392, wherein the antibody or antigen-bindingfragment thereof specifically binds to CUB domain-containing protein 1(CDCP1).

In some embodiments, the antibody or antigen-binding fragmentspecifically binds to human CDCP1 and/or Cynomolgus CDCP1. In someembodiments, the antibody or antigen-binding fragment has a dissociationconstant (K_(d)) for human CDCP1 that is less than 10 nM, and/or adissociation constant (K_(d)) for Cynomolgus CDCP1 that is less than 10nM. In some embodiments, the antibody or antigen-binding fragment is ahumanized antibody or antigen-binding fragment thereof.

In another aspect, the disclosure relates to an antibody orantigen-binding fragment thereof, wherein the antibody orantigen-binding fragment thereof binds to human CDCP1, and blockscleavage of human CDCP1 at residue 342. In some embodiments, theantibody or antigen-binding fragment thereof comprises a VH having CDRs1, 2, 3 with amino acid sequences set forth in SEQ ID NOs: 85, 86, and87, and a VL having CDRs 1, 2, 3 with amino acid sequences set forth inSEQ ID NOs: 88, 89, and 90. In some embodiments, the antibody orantigen-binding fragment thereof comprises a VH having CDRs 1, 2, 3 withamino acid sequences set forth in SEQ ID NOs: 95, 96, and 97, and a VLhaving CDRs 1, 2, 3 with amino acid sequences set forth in SEQ ID NOs:98, 99, and 100.

In some embodiments, the complementarity determining region (CDR)sequences in the heavy chain variable region and the light chainvariable region comprise or consist of the CDR sequences as set forth inFIG. 20 . In some embodiments, the CDR sequences in the heavy chainvariable region and the light chain variable region are at least 80%,90%, 92%, 95%, 97%, 98%, 99%, or 100% identical to the CDR sequences asset forth in FIG. 20 . In some embodiments, the CDR sequences in theheavy chain variable region and the light chain variable region differfrom the CDR sequences in FIG. 20 by one, two, three, four, or fiveamino acids. In some embodiments, the CDRs in FIG. 20 are substituted byone, two, three, four, or five conservative amino acids.

To generate and to select CDRs having preferred CDCP1 binding and/orneutralizing activity with respect to hCDCP1, standard methods known inthe art for generating antibodies, or antigen binding portions thereof,and assessing the CDCP1 binding and/or neutralizing characteristics ofthose antibodies, or antigen binding portions thereof, may be used,including but not limited to those specifically described herein.

In certain embodiments, the antibody comprises a heavy chain constantregion, such as an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgDconstant region. In certain embodiments, the anti-CDCP1 antibody, orantigen binding portion thereof, comprises a heavy chain immunoglobulinconstant domain selected from the group consisting of a human IgGconstant domain, a human IgM constant domain, a human IgE constantdomain, and a human IgA constant domain. In further embodiments, theantibody, or antigen binding portion thereof, has an IgG1 heavy chainconstant region, an IgG2 heavy chain constant region, an IgG3 constantregion, or an IgG4 heavy chain constant region. Preferably, the heavychain constant region is an IgG1 heavy chain constant region or an IgG4heavy chain constant region. Furthermore, the antibody can comprise alight chain constant region, either a kappa light chain constant regionor a lambda light chain constant region. Preferably, the antibodycomprises a kappa light chain constant region. Alternatively, theantibody portion can be, for example, a Fab fragment or a single chainFv fragment.

In certain embodiments, the anti-CDCP1 antibody binding portion is aFab, a Fab′, a F(ab′)2, a Fv, a disulfide linked Fv, an scFv, a singledomain antibody, or a diabody.

In certain embodiments, the anti-CDCP1 antibody, or antigen bindingportion thereof, is a multispecific antibody, e.g. a bispecificantibody.

Replacements of amino acid residues in the Fc portion to alter antibodyeffector function are have been described (Winter, et al. U.S. Pat. Nos.5,648,260 and 5,624,821, incorporated by reference herein). The Fcportion of an antibody mediates several important effector functionse.g. cytokine induction, ADCC, phagocytosis, complement dependentcytotoxicity (CDC) and half-life/clearance rate of antibody andantigen-antibody complexes. In some cases these effector functions aredesirable for therapeutic antibody but in other cases might beunnecessary or even deleterious, depending on the therapeuticobjectives. Certain human IgG isotypes, particularly IgG1 and IgG3,mediate ADCC and CDC via binding to Fc□Rs and complement C1q,respectively. Neonatal Fc receptors (FcRn) are the critical componentsdetermining the circulating half-life of antibodies. In still anotherembodiment at least one amino acid residue is replaced in the constantregion of the antibody, for example the Fc region of the antibody, suchthat effector functions of the antibody are altered.

One embodiment includes a labeled anti-CDCP1 antibody, or antibodyportion thereof, where the antibody is derivatized or linked to one ormore functional molecule(s) (e.g., another peptide or protein). Forexample, a labeled antibody can be derived by functionally linking anantibody or antibody portion of the disclosure (by chemical coupling,genetic fusion, noncovalent association or otherwise) to one or moreother molecular entities, such as another antibody (e.g., a bispecificantibody or a diabody), a detectable agent, a pharmaceutical agent, aprotein or peptide that can mediate the association of the antibody orantibody portion with another molecule (such as a streptavidin coreregion or a polyhistidine tag), and/or a cytotoxic or therapeutic agentselected from the group consisting of a mitotic inhibitor, an antitumorantibiotic, an immunomodulating agent, a vector for gene therapy, analkylating agent, an antiangiogenic agent, an antimetabolite, aboron-containing agent, a chemoprotective agent, a hormone, anantihormone agent, a corticosteroid, a photoactive therapeutic agent, anoligonucleotide, a radionuclide agent, a topoisomerase inhibitor, atyrosine kinase inhibitor, a radiosensitizer, and a combination thereof.

Useful detectable agents with which an antibody or antibody portionthereof, may be derivatized include fluorescent compounds. Exemplaryfluorescent detectable agents include fluorescein, fluoresceinisothiocyanate, rhodamine, 5-dimethylamine-1-napthalenesulfonylchloride, phycocrythrin and the like. An antibody may also bederivatized with detectable enzymes, such as alkaline phosphatase,horseradish peroxidase, glucose oxidase and the like. When an antibodyis derivatized with a detectable enzyme, it is detected by addingadditional reagents that the enzyme uses to produce a detectablereaction product. For example, when the detectable agent horseradishperoxidase is present the addition of hydrogen peroxide anddiaminobenzidine leads to a colored reaction product, which isdetectable. An antibody may also be derivatized with biotin, anddetected through indirect measurement of avidin or streptavidin binding.

In one embodiment, the antibody is conjugated to an imaging agent.Examples of imaging agents that may be used in the compositions andmethods described herein include, but are not limited to, a radiolabel(e.g., indium), an enzyme, a fluorescent label, a luminescent label, abioluminescent label, a magnetic label, and biotin.

In one embodiment, the antibodies or ADCs are linked to a radiolabel,such as, but not limited to, indium (¹¹¹In). ¹¹¹Indium may be used tolabel the antibodies and ADCs described herein for use in identifyingCDCP1 positive tumors. In a certain embodiment, anti-CDCP1 antibodies(or ADCs) described herein are labeled with ¹¹¹I via a bifunctionalchelator which is a bifunctional cyclohexyldiethylenetriaminepentaacetic acid (DTPA) chelate (see U.S. Pat. Nos.5,124,471; 5,434,287; and 5,286,850, each of which is incorporatedherein by reference).

Another embodiment of the disclosure provides a glycosylated bindingprotein wherein the anti-CDCP1 antibody or antigen binding portionthereof comprises one or more carbohydrate residues. Nascent in vivoprotein production may undergo further processing, known aspost-translational modification. In particular, sugar (glycosyl)residues may be added enzymatically, a process known as glycosylation.The resulting proteins bearing covalently linked oligosaccharide sidechains are known as glycosylated proteins or glycoproteins. Antibodiesare glycoproteins with one or more carbohydrate residues in the Fcdomain, as well as the variable domain. Carbohydrate residues in the Fcdomain have important effect on the effector function of the Fc domain,with minimal effect on antigen binding or half-life of the antibody (R.Jefferis, Biotechnol. Prog. 21 (2005), pp. 11-16). In contrast,glycosylation of the variable domain may have an effect on the antigenbinding activity of the antibody. Glycosylation in the variable domainmay have a negative effect on antibody binding affinity, likely due tosteric hindrance (Co, M. S., et al., Mol. Immunol. (1993) 30:1361-1367),or result in increased affinity for the antigen (Wallick, S. C., et al.,Exp. Med. (1988) 168:1099-1109; Wright, A., et al., EMBO J. (1991)10:2717-2723).

One aspect of the disclosure is directed to generating glycosylationsite mutants in which the O- or N-linked glycosylation site of thebinding protein has been mutated. One skilled in the art can generatesuch mutants using standard well-known technologies. Glycosylation sitemutants that retain the biological activity, but have increased ordecreased binding activity, are another object of the disclosure.

In still another embodiment, the glycosylation of the anti-CDCP1antibody or antigen binding portion is modified. For example, anaglycoslatcd antibody can be made (i.e., the antibody lacksglycosylation). Glycosylation can be altered to, for example, increasethe affinity of the antibody for antigen. Such carbohydratemodifications can be accomplished by, for example, altering one or moresites of glycosylation within the antibody sequence. For example, one ormore amino acid substitutions can be made that result in elimination ofone or more variable region glycosylation sites to thereby eliminateglycosylation at that site. Such aglycosylation may increase theaffinity of the antibody for antigen. Such an approach is described infurther detail in PCT Publication WO2003016466A2, and U.S. Pat. Nos.5,714,350 and 6,350,861, each of which is incorporated herein byreference in its entirety.

Additionally or alternatively, a modified anti-CDCP1 antibody can bemade that has an altered type of glycosylation, such as ahypofucosylated antibody having reduced amounts of fucosyl residues oran antibody having increased bisecting GlcNAc structures. Such alteredglycosylation patterns have been demonstrated to increase the ADCCability of antibodies. Such carbohydrate modifications can beaccomplished by, for example, expressing the antibody in a host cellwith altered glycosylation machinery. Cells with altered glycosylationmachinery have been described in the art and can be used as host cellsin which to express recombinant antibodies to thereby produce anantibody with altered glycosylation. See, for example, Shields, R. L. etal. (2002) J. Biol. Chem. 277:26733-26740; Umana et al. (1999) Nat.Biotech. 17:176-1, as well as, European Patent No: EP 1,176,195; PCTPublications WO 03/035835; WO 99/54342 80, each of which is incorporatedherein by reference in its entirety.

Protein glycosylation depends on the amino acid sequence of the proteinof interest, as well as the host cell in which the protein is expressed.Different organisms may produce different glycosylation enzymes (e.g.,glycosyltransferases and glycosidases), and have different substrates(nucleotide sugars) available. Due to such factors, proteinglycosylation pattern, and composition of glycosyl residues, may differdepending on the host system in which the particular protein isexpressed. Glycosyl residues useful may include, but are not limited to,glucose, galactose, mannose, fucose, n-acetylglucosamine and sialicacid. Preferably the glycosylated binding protein comprises glycosylresidues such that the glycosylation pattern is human.

Differing protein glycosylation may result in differing proteincharacteristics. For instance, the efficacy of a therapeutic proteinproduced in a microorganism host, such as yeast, and glycosylatedutilizing the yeast endogenous pathway may be reduced compared to thatof the same protein expressed in a mammalian cell, such as a CHO cellline. Such glycoproteins may also be immunogenic in humans and showreduced half-life in vivo after administration. Specific receptors inhumans and other animals may recognize specific glycosyl residues andpromote the rapid clearance of the protein from the bloodstream. Otheradverse effects may include changes in protein folding, solubility,susceptibility to proteases, trafficking, transport,compartmentalization, secretion, recognition by other proteins orfactors, antigenicity, or allergenicity. Accordingly, a practitioner mayprefer a therapeutic protein with a specific composition and pattern ofglycosylation, for example glycosylation composition and patternidentical, or at least similar, to that produced in human cells or inthe species-specific cells of the intended subject animal.

Expressing glycosylated proteins different from that of a host cell maybe achieved by genetically modifying the host cell to expressheterologous glycosylation enzymes. Using recombinant techniques, apractitioner may generate antibodies or antigen binding portions thereofexhibiting human protein glycosylation. For example, yeast strains havebeen genetically modified to express non-naturally occurringglycosylation enzymes such that glycosylated proteins (glycoproteins)produced in these yeast strains exhibit protein glycosylation identicalto that of animal cells, especially human cells (U.S. patent PublicationNos. 20040018590 and 20020137134 and PCT publication WO2005100584 A2).

Antibodies may be produced by any of a number of techniques. Forexample, expression from host cells, wherein expression vector(s)encoding the heavy and light chains is (are) transfected into a hostcell by standard techniques. The various forms of the term“transfection” are intended to encompass a wide variety of techniquescommonly used for the introduction of exogenous DNA into a prokaryoticor eukaryotic host cell, e.g., electroporation, calcium-phosphateprecipitation, DEAE-dextran transfection and the like. Although it ispossible to express antibodies in either prokaryotic or eukaryotic hostcells, expression of antibodies in eukaryotic cells is preferable, andmost preferable in mammalian host cells, because such eukaryotic cells(and in particular mammalian cells) are more likely than prokaryoticcells to assemble and secrete a properly folded and immunologicallyactive antibody.

Preferred mammalian host cells for expressing the recombinant antibodiesdisclosed herein include Chinese Hamster Ovary (CHO cells) (includingdhfr-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad.So. USA 77:4216-4220, used with a DHFR selectable marker, e.g., asdescribed in R. J. Kaufman and P. A. Sharp (1982) Mol. Biol.159:601-621), NS0 myeloma cells, COS cells and SP2 cells. Whenrecombinant expression vectors encoding antibody genes are introducedinto mammalian host cells, the antibodies are produced by culturing thehost cells for a period of time sufficient to allow for expression ofthe antibody in the host cells or, more preferably, secretion of theantibody into the culture medium in which the host cells are grown.Antibodies can be recovered from the culture medium using standardprotein purification methods.

Host cells can also be used to produce functional antibody fragments,such as Fab fragments or scFv molecules. It will be understood thatvariations on the above procedure are within the scope of thedisclosure. For example, it may be desirable to transfect a host cellwith DNA encoding functional fragments of either the light chain and/orthe heavy chain of an antibody. Recombinant DNA technology may also beused to remove some, or all, of the DNA encoding either or both of thelight and heavy chains that is not necessary for binding to the antigensof interest. The molecules expressed from such truncated DNA moleculesare also encompassed by the antibodies of the disclosure. In addition,bifunctional antibodies may be produced in which one heavy and one lightchain are an antibody of the disclosure and the other heavy and lightchain are specific for an antigen other than the antigens of interest bycrosslinking an antibody of the disclosure to a second antibody bystandard chemical crosslinking methods.

In a preferred system for recombinant expression of an antibody, orantigen binding portion thereof, a recombinant expression vectorencoding both the antibody heavy chain and the antibody light chain isintroduced into CHO cells comprising a glutamine synthase expressionsystem, commercially available from Lonza (hereafter GS-CHO)(Bebbington, C. R. et al. (1992), Biotechnology, 10, pages 169-175).

In another system for recombinant expression of an antibody, or antigenbinding portion thereof, a recombinant expression vector encoding boththe antibody heavy chain and the antibody light chain is introduced intodhfr-CHO cells by calcium phosphate-mediated transfection. Within therecombinant expression vector, the antibody heavy and light chain genesare each operatively linked to CMV enhancer/AdMLP promoter regulatoryelements to drive high levels of transcription of the genes. Therecombinant expression vector also carries a DHFR gene, which allows forselection of CHO cells that have been transfected with the vector usingmethotrexate selection/amplification. The selected transformant hostcells are cultured to allow for expression of the antibody heavy andlight chains and intact antibody is recovered from the culture medium.Standard molecular biology techniques are used to prepare therecombinant expression vector, transfcct the host cells, select fortransformants, culture the host cells and recover the antibody from theculture medium. Still further the disclosure provides a method ofsynthesizing a recombinant antibody by culturing a host cell in asuitable culture medium until a recombinant antibody is synthesized.Recombinant antibodies may be produced using nucleic acid moleculescorresponding to the amino acid sequences disclosed herein. In oneembodiment, the nucleic acid molecules set forth in SEQ ID NOs: 82-101are used in the production of a recombinant antibody. The method canfurther comprise isolating the recombinant antibody from the culturemedium.

III. Anti-CDCP1 Antibody Drug Conjugates (ADCs)

Anti-CDCP1 antibodies described herein may be conjugated to a drugmoiety to form an anti-CDCP1 Antibody Drug Conjugate (ADC).Antibody-drug conjugates (ADCs) may increase the therapeutic efficacy ofantibodies in treating disease, e.g., cancer, due to the ability of theADC to selectively deliver one or more drug moiety(s) to target tissuesor cells, e.g., CDCP1 expressing tumors or CDCP1 expressing cells. Thus,in certain embodiments, the disclosure provides anti-CDCP1 ADCs fortherapeutic use, e.g., treatment of cancer.

Anti-CDCP1 ADCs comprise an anti-CDCP1 antibody, i.e., an antibody thatspecifically binds to CDCP1, linked to one or more drug moieties. Thespecificity of the ADC is defined by the specificity of the antibody,i.e., anti-CDCP1. In one embodiment, an anti-CDCP1 antibody is linked toone or more cytotoxic drug(s) which is delivered internally to a cancercell expressing CDCP1.

Examples of drugs that may be used in the anti-CDCP1 ADCs are providedbelow, as are linkers that may be used to conjugate the antibody and theone or more drug(s). The terms “drug,” “agent,” and “drug moiety” areused interchangeably herein. The terms “linked” and “conjugated” arealso used interchangeably herein and indicate that the antibody andmoiety are covalently linked.

In some embodiments, the ADC has the following formula (formula I):Ab-(L-D)_(n)  (I)wherein Ab an anti-CDCP1 antibody described herein, and (L-D) is aLinker-Drug moiety. The Linker-Drug moiety is made of L- which is aLinker, and -D, which is a drug moiety having, for example, cytostatic,cytotoxic, or otherwise therapeutic activity against a target cell,e.g., a cell expressing CDCP1; and n is an integer from 1 to 20. In someembodiments, n ranges from 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to3, 1 to 2, or is 1. The DAR of an ADC is equivalent to the “n” referredto in Formula I.

Additional details regarding drugs (D of Formula I) and linkers (L ofFormula I) that may be used in the ADCs, as well as alternative ADCstructures, are described below.

A. Anti-CDCP1 ADCs: Exemplary Drugs for Conjugation

Anti-CDCP1 antibodies may be used in ADCs to target one or more drug(s)to a cell of interest, e.g., a cell expressing CDCP1. The anti-CDCP1ADCs disclosed herein provide a targeted therapy that may, for example,reduce the side effects often seen with anti-cancer therapies, as theone or more drug(s) is delivered to a specific cell. In one embodiment,the drug used in an ADC is saporin. In another embodiment, the drug usedin an ADC is dacarbazine. In another embodiment, the drug used in an ADCis carboplatin.

Examples of drugs that may be used in ADCs, i.e., drugs that may beconjugated to the anti-CDCP1 antibodies, are provided below, and includemitotic inhibitors, antitumor antibiotics, immunomodulating agents, genetherapy vectors, alkylating agents, antiangiogenic agents,antimetabolites, boron-containing agents, chemoprotective agents,hormone agents, glucocorticoids, photoactive therapeutic agents,oligonucleotides, radioactive isotopes, radiosensitizers, topoisomeraseinhibitors, tyrosine kinase inhibitors, and combinations thereof.

I. Mitotic Inhibitors

In one aspect, anti-CDCP1 antibodies may be conjugated to one or moremitotic inhibitor(s) to form an ADC for the treatment of cancer. Theterm “mitotic inhibitor”, as used herein, refers to a cytotoxic and/ortherapeutic agent that blocks mitosis or cell division, a biologicalprocess particularly important to cancer cells. A mitotic inhibitordisrupts microtubules such that cell division is prevented, often byeffecting microtubule polymerization (e.g., inhibiting microtubulepolymerization) or microtubule depolymerization (e.g., stabilizing themicrotubule cytoskeleton against depolymrization). Thus, in oneembodiment, an anti-CDCP1 antibody of the invention is conjugated to oneor more mitotic inhibitor(s) that disrupts microtubule formation byinhibiting tubulin polymerization. In another embodiment, an anti-CDCP1antibody of the invention is conjugated to one or more mitoticinhibitor(s) that stabilizes the microtubule cytoskeleton fromdeploymerization. In one embodiment, the mitotic inhibitor used in theADCs of the invention is Ixempra (ixabepilone). Examples of mitoticinhibitors that may be used in the anti-CDCP1 ADCs of the invention areprovided below. Included in the genus of mitotic inhibitors areauristatins, described below.

a. Dolastatins

The anti-CDCP1 antibodies of the invention may be conjugated to at leastone dolastatin to form an ADC. Dolastatins are short peptidic compoundsisolated from the Indian Ocean sea hare Dolabella auricularia (seePettit et al., J. Am. Chem. Soc., 1976, 98, 4677). Examples ofdolastatins include dolastatin 10 and dolatstin 15. Dolastatin 15, aseven-subunit depsipeptide derived from Dolabella auricularia, and is apotent antimitotic agent structurally related to the antitubulin agentdolastatin 10, a five-subunit peptide obtained from the same organism.Thus, in one embodiment, the anti-CDCP1 ADC of the invention comprisesan anti-CDCP1 antibody, as described herein, and at least onedolastatin. Auristatins are synthetic derivatives of dolastatin 10.

b. Auristatins

Anti-CDCP1 antibodies may be conjugated to at least one auristatin.Auristatins represent a group of dolastatin analogs that have generallybeen shown to possess anticancer activity by interfering withmicrotubule dynamics and GTP hydrolysis, thereby inhibiting cellulardivision. For example, Auristatin E (U.S. Pat. No. 5,635,483) is asynthetic analogue of the marine natural product dolastatin 10, acompound that inhibits tubulin polymerization by binding to the samesite on tubulin as the anticancer drug vincristine (G. R. Pettit, Prog.Chem. Org. Nat. Prod, 70: 1-79 (1997)). Dolastatin 10, auristatin PE,and auristatin E are linear peptides having four amino acids, three ofwhich are unique to the dolastatin class of compounds. Exemplaryembodiments of the auristatin subclass of mitotic inhibitors include,but are not limited to, monomethyl auristatin D (MMAD or auristatin Dderivative), monomethyl auristatin E (MMAE or auristatin E derivative),monomethyl auristatin F (MMAF or auristatin F derivative), auristatin Fphenylenediamine (AFP), auristatin EB (AEB), auristatin EFP (AEFP), and5-benzoylvaleric acid-AE ester (AEVB). The synthesis and structure ofauristatin derivatives are described in U.S. Patent ApplicationPublication Nos. 2003-0083263, 2005-0238649 and 2005-0009751;International Patent Publication No. WO 04/010957, International PatentPublication No. WO 02/088172, and U.S. Pat. Nos. 6,323,315; 6,239,104;6,034,065; 5,780,588; 5,665,860; 5,663,149; 5,635,483; 5,599,902;5,554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036;5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,816,444; and 4,486,414,each of which is incorporated by reference herein.

In one embodiment, anti-CDCP1 antibodies are conjugated to at least oneMMAE (mono-methyl auristatin E). Monomethyl auristatin E (MMAE, vedotin)inhibits cell division by blocking the polymerization of tubulin.Because of its super toxicity, it also cannot be used as a drug itself.In recent cancer therapy developments, it is linked to a monoclonalantibody (mAb) that recognizes a specific marker expression in cancercells and directs MMAE to the cancer cells. In one embodiment, thelinker linking MMAE to the anti-CDCP1 antibody is stable inextracellular fluid (i.e., the medium or environment that is external tocells), but is cleaved by cathepsin once the ADC has bound to thespecific cancer cell antigen and entered the cancer cell, thus releasingthe toxic MMAE and activating the potent anti-mitotic mechanism.

The structure of MMAE is provided below.

In one embodiment, the antibody is coupled to a single drug and,therefore, has a DAR of 1. In certain embodiments, the ADC will have aDAR of 2 to 8, or, alternatively, 2 to 4.

c. Maytansinoids

The anti-CDCP1 antibodies of the invention may be conjugated to at leastone maytansinoid to form an ADC. Maytansinoids are potent antitumoragents that were originally isolated from members of the higher plantfamilies Celastraceae, Rhamnaceae, and Euphorbiaceae, as well as somespecies of mosses (Kupchan et al, J. Am. Chem. Soc. 94:1354-1356 [1972];Wani et al, J. Chem. Soc. Chem. Commun. 390: [1973]; Powell et al, J.Nat. Prod. 46:660-666 [1983]; Sakai et al, J. Nat. Prod. 51:845-850[1988]; and Suwanborirux et al, Experientia 46:117-120 [1990]). Evidencesuggests that maytansinoids inhibit mitosis by inhibiting polymerizationof the microtubule protein tubulin, thereby preventing formation ofmicrotubules (see, e.g., U.S. Pat. No. 6,441,163 and Remillard et al.,Science, 189, 1002-1005 (1975)). Maytansinoids have been shown toinhibit tumor cell growth in vitro using cell culture models, and invivo using laboratory animal systems. Moreover, the cytotoxicity ofmaytansinoids is 1,000-fold greater than conventional chemotherapeuticagents, such as, for example, methotrexate, daunorubicin, andvincristine (see, e.g., U.S. Pat. No. 5,208,020).

Maytansinoids to include maytansine, maytansinol, C-3 esters ofmaytansinol, and other maytansinol analogues and derivatives (see, e.g.,U.S. Pat. Nos. 5,208,020 and 6,441,163, each of which is incorporated byreference herein). C-3 esters of maytansinol can be naturally occurringor synthetically derived. Moreover, both naturally occurring andsynthetic C-3 maytansinol esters can be classified as a C-3 ester withsimple carboxylic acids, or a C-3 ester with derivatives ofN-methyl-L-alanine, the latter being more cytotoxic than the former.Synthetic maytansinoid analogues are described in, for example, Kupchanet al., J. Med. Chem., 21, 31-37 (1978).

Suitable maytansinoids for use in ADCs of the invention can be isolatedfrom natural sources, synthetically produced, or semi-syntheticallyproduced. Moreover, the maytansinoid can be modified in any suitablemanner, so long as sufficient cytotoxicity is preserved in the ultimateconjugate molecule. In this regard, maytansinoids lack suitablefunctional groups to which antibodies can be linked. A linking moietydesirably is utilized to link the maytansinoid to the antibody to formthe conjugate, and is described in more detail in the linker sectionbelow. The structure of an exemplary maytansinoid, mertansine (DM1), isprovided below.

Representative examples of maytansinoids include, but are not limited,to DM1 (N^(2′)-deacetyl-N^(2′)-(3-mercapto-1-oxopropyl)-maytansine; alsoreferred to as mertansine, drug maytansinoid 1; ImmunoGen, Inc.; seealso Chari et al. (1992) Cancer Res 52:127), DM2, DM3(N^(2′)-deacetyl-N^(2′)-(4-mercapto-1-oxopentyl)-maytansine), DM4(4-methyl-4-mercapto-1-oxopentyl)-maytansine), and maytansinol (asynthetic maytansinoid analog). Other examples of maytansinoids aredescribed in U.S. Pat. No. 8,142,784, incorporated by reference herein.

Ansamitocins are a group of maytansinoid antibiotics that have beenisolated from various bacterial sources. These compounds have potentantitumor activities. Representative examples include, but are notlimited to ansamitocin P1, ansamitocin P2, ansamitocin P3, andansamitocin P4.

In one embodiment of the invention, an anti-CDCP1 antibody is conjugatedto at least one DM1. In one embodiment, an anti-CDCP1 antibody isconjugated to at least one DM2. In one embodiment, an anti-CDCP1antibody is conjugated to at least one DM3. In one embodiment, ananti-CDCP1 antibody is conjugated to at least one DM4.

2. Antitumor Antibiotics

Anti-CDCP1 antibodies may be conjugated to one or more antitumorantibiotic(s) for the treatment of cancer. As used herein, the term“antitumor antibiotic” means an antineoplastic drug that blocks cellgrowth by interfering with DNA and is made from a microorganism. Often,antitumor antibiotics either break up DNA strands or slow down or stopDNA synthesis. Examples of antitumor antibiotics that may be included inthe anti-CDCP1 ADCs include, but are not limited to, actinomycines(e.g., pynolo[2,1-c][1.4]benzodiazepines), anthracyclines,calicheamicins, and duocarmycins. In addition to the foregoing,additional antitumor antibiotics that may be used in the anti-CDCP1 ADCsinclude bleomycin (Blenoxane, Bristol-Myers Squibb), mitomycin, andplicamycin (also known as mithramycin).

3. Immunomodulating Agents

In one aspect, anti-CDCP1 antibodies may be conjugated to at least oneimmunomodulating agent. As used herein, the term “immunomodulatingagent” refers to an agent that can stimulate or modify an immuneresponse. In one embodiment, an immunomodulating agent is animmunostimuator which enhances a subject's immune response. In anotherembodiment, an immunomodulating agent is an immunosuppressant whichprevents or decreases a subject's immune response. An immunomodulatingagent may modulate myeloid cells (monocytes, macrophages, dendriticcells, megakaryocytes and granulocytes) or lymphoid cells (T cells, Bcells and natural killer (NK) cells) and any further differentiated cellthereof. Representative examples include, but are not limited to,bacillus calmette-gucrin (BCG) and levamisole (Ergamisol). Otherexamples of immunomodulating agents that may be used in the ADCsinclude, but are not limited to, cancer vaccines, and cytokines.

As used herein, the term “cancer vaccine” refers to a composition (e.g.,a tumor antigen and a cytokine) that elicits a tumor-specific immuneresponse. The response is elicited from the subject's own immune systemby administering the cancer vaccine, or, in the case of the instantdisclosure, administering an ADC comprising an anti-CDCP1 antibody and acancer vaccine. In preferred embodiments, the immune response results inthe eradication of tumor cells in the body (e.g., primary or metastatictumor cells). The use of cancer vaccines generally involves theadministration of a particular antigen or group of antigens that are,for example, present on the surface a particular cancer cell, or presenton the surface of a particular infectious agent shown to facilitatecancer formation. In some embodiments, the use of cancer vaccines is forprophylactic purposes, while in other embodiments, the use is fortherapeutic purposes. Non-limiting examples of cancer vaccines that maybe used in the anti-CDCP1 ADCs include, recombinant bivalent humanpapillomavirus (HPV) vaccine types 16 and 18 vaccine (Cervarix,GlaxoSmithKline), recombinant quadrivalent human papillomavirus (HPV)types 6, 11, 16, and 18 vaccine (Gardasil, Merck & Company), andsipuleucel-T (Provenge, Dendreon). Thus, in one embodiment, theanti-CDCP1 antibody is conjugated to at least one cancer vaccine that iseither an immunostimulator or is an immunosuppressant.

The anti-CDCP1 antibodies may be conjugated to at least one cytokine.The term “cytokine” generally refers to proteins released by one cellpopulation which act on another cell as intercellular mediators.Cytokines directly stimulate immune effector cells and stromal cells atthe tumor site and enhance tumor cell recognition by cytotoxic effectorcells (Lee and Margolin (2011) Cancers 3:3856). Numerous animal tumormodel studies have demonstrated that cytokines have broad anti-tumoractivity and this has been translated into a number of cytokine-basedapproaches for cancer therapy (Lee and Margoli, supra). Recent yearshave seen a number of cytokines, including GM-CSF, IL-7, IL-12, IL-15,IL-18 and IL-21, enter clinical trials for patients with advanced cancer(Lee and Margoli, supra).

Examples of cytokines that may be used in the ADCs include, but are notlimited to, parathyroid hormone; thyroxine; insulin; proinsulin;relaxin; prorelaxin; glycoprotein hormones such as follicle stimulatinghormone (FSH), thyroid stimulating hormone (TSH), and luteinizinghormone (LH); hepatic growth factor; fibroblast growth factor;prolactin; placental lactogen; tumor necrosis factor;mullerian-inhibiting substance; mouse gonadotropin-associated peptide;inhibin; activin; vascular endothelial growth factor; integrin;thrombopoietin (TPO); nerve growth factors such as NGF; platelet-growthfactor; transforming growth factors (TGFs); insulin-like growth factor-Iand -II; erythropoietin (EPO); osteoinductive factors; interferons suchas interferon α, β, and γ, colony stimulating factors (CSFs);granulocyte-macrophage-C-SF (GM-CSF); and granulocyte-CSF (G-CSF);interleukins (ILs) such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6,IL-7, IL-8, IL-9, IL-11, IL-12; tumor necrosis factor; and otherpolypeptide factors including LIF and kit ligand (KL). As used herein,the term cytokine includes proteins from natural sources or fromrecombinant cell culture and biologically active equivalents of thenative sequence cytokines. Thus, in one embodiment, the disclosureprovides an ADC comprising an anti-CDCP1 antibody described herein and acytokine.

The anti-CDCP1 antibodies may be conjugated to at least one colonystimulating factor (CSF). Colony stimulating factors (CSFs) are growthfactors that assist the bone marrow in making red blood cells. Becausesome cancer treatments (e.g., chemotherapy) can affect white blood cells(which help fight infection), colony-stimulating factors may beintroduced to help support white blood cell levels and strengthen theimmune system. Colony-stimulating factors may also be used following abone marrow transplant to help the new marrow start producing whiteblood cells. Representative examples of CSFs that may be used in theanti-CDCP1 ADCs include, but are not limited to erythropoietin(Epoetin), filgrastim (Neopogen (also known as granulocytecolony-stimulating factor (G-CSF); Amgen, Inc.), sargramostim (leukine(granulocyte-macrophage colony-stimulating factor and GM-CSF); GenzymeCorporation), promegapoietin, and Oprelvekin (recombinant IL-11; Pfizer,Inc.). Thus, in one embodiment, an ADC may comprise an anti-CDCP1antibody described herein and a CSF.

4. Alkylating Agents

The anti-CDCP1 antibodies may be conjugated to one or more alkylatingagent(s). Alkylating agents are a class of antineoplastic compounds thatattaches an alkyl group to DNA. Examples of alkylating agents that maybe used in the ADCs include, but are not limited to, alkyl sulfonates,ethylenimimes, methylamine derivatives, epoxides, nitrogen mustards,nitrosoureas, triazines and hydrazines.

DNA Alkylating Agents

The term “DNA alkylating agent”, as used herein, includes a family ofDNA alkylating agents including indolino-benzodiazepines (IGNs). IGNsrepresent a chemical class of cytotoxic molecules with high in vitropotency (IC₅₀ values in the low pmol/L range) toward cancer cells.Examples of IGN DNA alkylating agents that can be used as a cytotoxicpayload in an ADC are described in Miller et al. (2016) Molecular CancerTherapeutics, 15(8)). The IGN compounds described in Miller et al. bindto the minor groove of DNA followed by covalent reaction of guanineresidues with the two imine functionalities in the molecule resulting incross-linking of DNA. The structure of an exemplary IGN is providedbelow.

5. Antiangiogenic Agents

In one aspect, the anti-CDCP1 antibodies described herein are conjugatedto at least one antiangiogenic agent. Antiangiogenic agents inhibit thegrowth of new blood vessels. Antiangiogenic agents exert their effectsin a variety of ways. In some embodiments, these agents interfere withthe ability of a growth factor to reach its target. For example,vascular endothelial growth factor (VEGF) is one of the primary proteinsinvolved in initiating angiogenesis by binding to particular receptorson a cell surface. Thus, certain antiangiogenic agents, that prevent theinteraction of VEGF with its cognate receptor, prevent VEGF frominitiating angiogenesis. In other embodiments, these agents interferewith intracellular signaling cascades. For example, once a particularreceptor on a cell surface has been triggered, a cascade of otherchemical signals is initiated to promote the growth of blood vessels.Thus, certain enzymes, for example, some tyrosine kinases, that areknown to facilitate intracellular signaling cascades that contribute to,for example, cell proliferation, are targets for cancer treatment. Inother embodiments, these agents interfere with intercellular signalingcascades. Yet, in other embodiments, these agents disable specifictargets that activate and promote cell growth or by directly interferingwith the growth of blood vessel cells. Angiogenesis inhibitoryproperties have been discovered in more than 300 substances withnumerous direct and indirect inhibitory effects.

Representative examples of antiangiogenic agents that may be used in theADCs include, but are not limited to, angiostatin, ABX EGF, C1-1033,PKI-166, EGF vaccine, EKB-569, GW2016, ICR-62, EMD 55900, CP358,PD153035, AG1478, IMC-C225 (Erbitux, ZD1839 (Iressa), OSI-774, Erlotinib(tarceva), angiostatin, arrestin, endostatin, BAY 12-9566 andw/fluorouracil or doxorubicin, canstatin, carboxyamidotriozolc and withpaclitaxel, EMD121974, S-24, vitaxin, dimethylxanthenone acetic acid,IM862, Interleukin-12, Interleukin-2, NM-3, HuMV833, PTK787, RhuMab,angiozyme (ribozyme), IMC-1C11, Neovastat, marimstat, prinomastat,BMS-275291, COL-3, MM1270, SU101, SU6668, SU11248, SU5416, withpaclitaxel, with gemcitabine and cisplatin, and with irinotecan andcisplatin and with radiation, tecogalan, temozolomide and PEG interferonα2b, tetrathiomolybdate, TNP-470, thalidomide, CC-5013 and withtaxotcrc, tumstatin, 2-methoxyestradiol, VEGF trap, mTOR inhibitors(deforolimus, everolimus (Afinitor, Novartis PharmaceuticalCorporation), and temsirolimus (Torisel, Pfizer, Inc.)), tyrosine kinaseinhibitors (e.g., erlotinib (Tarceva, Genentech, Inc.), imatinib(Gleevec, Novartis Pharmaceutical Corporation), gefitinib (Iressa,AstraZeneca Pharmaceuticals), dasatinib (Sprycel, Brystol-Myers Squibb),sunitinib (Sutent, Pfizer, Inc.), nilotinib (Tasigna, NovartisPharmaceutical Corporation), lapatinib (Tykcrb, GlaxoSmithKlincPharmaceuticals), sorafenib (Nexavar, Bayer and Onyx), phosphoinositide3-kinases (PI3K).

6. Antimetabolites

The anti-CDCP1 antibodies may be conjugated to at least oneantimetabolite. Antimetabolites are types of chemotherapy treatmentsthat are very similar to normal substances within the cell. When thecells incorporate an antimetabolite into the cellular metabolism, theresult is negative for the cell, e.g., the cell is unable to divide.Antimetabolites are classified according to the substances with whichthey interfere. Examples of antimetabolies that may be used in the ADCsinclude, but are not limited to, a folic acid antagonist (e.g.,methotrexate), a pyrimidine antagonist (e.g., 5-Fluorouracil,Foxuridine, Cytarabine, Capecitabine, and Gemcitabine), a purineantagonist (e.g., 6-Mercaptopurine and 6-Thioguanine) and an adenosinedeaminase inhibitor (e.g., Cladribine, Fludarabine, Nelarabine andPentostatin), as described in more detail below.

7. Boron-Containing Agents

The anti-CDCP1 antibody may be conjugated to at least one boroncontaining agent. Boron-containing agents comprise a class of cancertherapeutic compounds which interfere with cell proliferation.Representative examples of boron containing agents include, but are notlimited, to borophycin and bortezomib (Velcade, MilleniumPharmaceuticals).

8. Chemoprotective Agents

The anti-CDCP1 antibodies may be conjugated to at least onechemoprotective agent. Chemoprotective drugs are a class of compounds,which help protect the body against specific toxic effects ofchemotherapy. Chemoprotective agents may be administered with variouschemotherapies in order to protect healthy cells from the toxic effectsof chemotherapy drugs, while simultaneously allowing the cancer cells tobe treated with the administered chemotherapeutic. Representativechemoprotective agents include, but are not limited to amifostine(Ethyol, Mcdimmunc, Inc.), which is used to reduce renal toxicityassociated with cumulative doses of cisplatin, dexrazoxane (Totect,Apricus Pharma; Zinecard), for the treatment of extravasation caused bythe administration of anthracycline (Totect), and for the treatment ofcardiac-related complications caused by the administration of theantitumor antibiotic doxorubicin (Zinecard), and mesna (Mesnex,Bristol-Myers Squibb), which is used to prevent hemorrhagic cystitisduring chemotherapy treatment with ifocfamide.

9. Photoactive Therapeutic Agents

The anti-CDCP1 antibodies may be conjugated to at least one photoactivetherapeutic agent. Photoactive therapeutic agents include compounds thatcan be deployed to kill treated cells upon exposure to electromagneticradiation of a particular wavelength. Therapeutically relevant compoundsabsorb electromagnetic radiation at wavelengths which penetrate tissue.In preferred embodiments, the compound is administered in a non-toxicform that is capable of producing a photochemical effect that is toxicto cells or tissue upon sufficient activation. In other preferredembodiments, these compounds are retained by cancerous tissue and arereadily cleared from normal tissues. Non-limiting examples includevarious chromagens and dyes.

10. Radionuclide Agents (Radioactive Isotopes)

The anti-CDCP1 antibodies may be conjugated to at least one radionuclideagent. Radionuclide agents comprise agents that are characterized by anunstable nucleus that is capable of undergoing radioactive decay. Thebasis for successful radionuclide treatment depends on sufficientconcentration and prolonged retention of the radionuclide by the cancercell. Other factors to consider include the radionuclide half-life, theenergy of the emitted particles, and the maximum range that the emittedparticle can travel. In preferred embodiments, the therapeutic agent isa radionuclide selected from the group consisting of ¹¹¹In, ¹⁷⁷Ln,²¹²Bi, ²¹³Bi, ²¹¹At, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁹⁰Y, ¹²⁵I, ¹³¹I, ³²P, ³³P, ⁴⁷Sc,¹¹¹Ag, ⁶⁷Ga, ¹⁴²Pr, ¹⁵³Sm, ¹⁶¹Tb, ¹⁶⁶Dy, ¹⁶⁶Ho, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁸⁹Re,²¹²Pb, ²²³Ra, ²²⁵Ac, ⁵⁹Fe, ⁷⁵Se, ⁷⁷As, ⁸⁹Sr, ⁹⁹Mo, ¹⁰⁵Rh, ¹⁰⁹Pd, ¹⁴³Pr,¹⁴⁹Pm, ¹⁶⁹Er, ¹⁹⁴Ir, ¹⁹⁸Au, ¹⁹⁹Au, an ²¹¹Pb. Also preferred areradionuclides that substantially decay with Auger-emitting particles.For example, Co-58, Ga-67, Br-80m, Tc-99m, Rh-103m, Pt-109, 1, Sb-119,1-125, Ho-161, Os-189m and Ir-192. Decay energies of usefulbeta-particle-emitting nuclides are preferably Dy-152, At-211, Bi-212,Ra-223, Rn-219, Po-215, Bi-21 1, Ac-225, Fr-221, At-217, Bi-213 andFm-255. Decay energies of useful alpha-particle-emitting radionuclidesare preferably 2,000-10,000 kcV, more preferably 3,000-8,000 keV, andmost preferably 4,000-7,000 keV. Additional potential radioisotopes ofuse include ¹¹C, ¹³N, ¹⁵0, ⁷⁵Br, ¹⁹⁸Au, ²²⁴AC, ¹²⁶I, ¹³³I, ⁷⁷Br,^(113m)In, ⁹⁵Ru, ⁹⁷Ru, ¹⁹³Ru, ¹⁹⁵Ru, ¹⁰⁷Hg, ²⁰³Hg, ^(121m)Te, ^(122m)Te,^(125m)Te, ¹⁶⁵Tm, ¹⁶⁷Tm, ¹⁶⁸Tm, ¹⁹⁷Pt, ¹⁰⁹Pd, ¹⁰⁵Rh, ¹⁴²Pr, ¹⁴³Pr,¹⁶¹Tb, ¹⁶⁶Ho, ¹⁹⁹Au, ¹⁹⁹Au, ⁵⁷Co, ⁵¹Cr, ⁵⁹Fe, ⁷⁵Se, ²⁰¹Tl, ²²⁵Ac, ⁷⁶Br,¹⁶⁹Yb, and the like.

11. Radiosensitizers

The anti-CDCP1 antibodies may be conjugated to at least oneradiosensitizer. The term “radiosensitizer,” as used herein, is definedas a molecule, preferably a low molecular weight molecule, administeredto animals in therapeutically effective amounts to increase thesensitivity of the cells to be radiosensitized to electromagneticradiation and/or to promote the treatment of diseases that are treatablewith electromagnetic radiation. Radiosensitizers are agents that makecancer cells more sensitive to radiation therapy, while typically havingmuch less of an effect on normal cells. Thus, the radiosensitizer can beused in combination with a radiolabeled antibody or ADC. The addition ofthe radiosensitizer can result in enhanced efficacy when compared totreatment with the radiolabeled antibody or antibody fragment alone.Radiosensitizers are described in D. M. Goldberg (ed.), Cancer Therapywith Radiolabeled Antibodies, CRC Press (1995). Examples ofradiosensitizers include gemcitabine, 5-fluorouracil, taxane, andcisplatin.

Radiosensitizers may be activated by the electromagnetic radiation ofX-rays. Representative examples of X-ray activated radiosensitizersinclude, but are not limited to, the following: metronidazole,misonidazole, desmethylmisonidazole, pimonidazole, etanidazole,nimorazole, mitomycin C, RSU 1069, SR 4233, E09, RB 6145, nicotinamide,5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR),bromodeoxycytidine, fluorodeoxyuridine (FUdR), hydroxyurea, cisplatin,and therapeutically effective analogs and derivatives of the same.Alternatively, radiosensitizers may be activated using photodynamictherapy (PDT). Representative examples of photodynamic radiosensitizersinclude, but are not limited to, hematoporphyrin derivatives,Photofrin(r), benzoporphyrin derivatives, NPe6, tin etioporphyrin(SnET2), pheoborbide a, bacteriochlorophyll a, naphthalocyanincs,phthalocyanines, zinc phthalocyanine, and therapeutically effectiveanalogs and derivatives of the same.

12. Topoisomerase Inhibitors

The anti-CDCP1 antibodies may be conjugated to at least onetopoisomerase inhibitor. Topoisomerase inhibitors are chemotherapyagents designed to interfere with the action of topoisomerase enzymes(topoisomerase I and II), which are enzymes that control the changes inDNA structure by catalyzing then breaking and rejoining of thephosphodiester backbone of DNA strands during the normal cell cycle.Representative examples of DNA topoisomerase I inhibitors include, butare not limited to, camptothecins and its derivatives irinotecan(CPT-11, Camptosar, Pfizer, Inc.) and topotecan (Hycamtin,GlaxoSmithKline Pharmaceuticals). Representative examples of DNAtopoisomerase II inhibitors include, but are not limited to, amsacrine,daunorubicin, doxotrubicin, epipodophyllotoxins, ellipticines,epirubicin, etoposide, razoxane, and teniposide.

13. Tyrosine Kinase Inhibitors

The anti-CDCP1 antibodies may be conjugated to at least one tyrosinekinase inhibitor. Tyrosine kinases are enzymes within the cell thatfunction to attach phosphate groups to the amino acid tyrosine. Byblocking the ability of protein tyrosine kinases to function, tumorgrowth may be inhibited. Examples of tyrosine kinases that may be usedon the ADCs include, but are not limited to, Axitinib, Bosutinib,Cediranib, Dasatinib, Erlotinib, Gefitinib, Imatinib, Lapatinib,Lestaurtinib, Nilotinib, Semaxanib, Sunitinib, and Vandetanib.

14. Additional Agents

Many other types of agents are well known to one of ordinary skill inthe art, and the anti-CDCP1 antibodies may be conjugated to any agentavailable to one of ordinary skill in the art. Agents include, but arenot limited to, cytotoxic agents, cytostatic agents, anti-angiogenicagents, debulking agents, chemotherapeutic agents, radiotherapy andradiotherapeutic agents, targeted anti-cancer agents, biologicalresponse modifiers, therapeutic antibodies, cancer vaccines, cytokines,hormone therapies, radiation therapy and anti-metastatic agents, andimmunotherapeutic agents.

In some implementations, the therapeutic agent is an anti-cancer agent.Examples of anti-cancer agents include, but are not limited to,cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin,etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin,daunorubicin, dihydroxy anthracin, dione, mitoxantrone, mithramycin,actinomycin D, 1-dchydrotestosterone, glucocorticoids, procaine,tetracaine, lidocaine, propranolol, puromycin, epirubicin, andcyclophosphamide and analogs or homologs thereof.

In other implementations, the therapeutic agent is a maytansinoidcell-killing agent, e.g.,N^(2′)-deacetyl-N^(2′)-(3-mercapto-1-oxopropyl)-maytansine (DM1) orN^(2′)-deacetyl-N^(2′)-(4-mercapto-4-methyl-1-oxopentyl) maytansine(DM4). DM1 is a sulfhydryl-containing derivative of maytansine that canbe linked to the peptide, e.g., via a disulfide linker that releases DM1when inside target cells. DM1 attached to an antibody with a thioetherlinker is called “emtansine” in its INN name (e.g., ado-trastuzumabemtansine). DM1 attached to an antibody with the N-succinimidyl4-(2-pyridyldithio)pentanoate (SPP) linker is called “mertansine.” DM4attached with the N-succinimidyl 3-(2-pyridyldithio)butyrate (SPDB)linker is called “ravtansinc,” e.g., indatuximab ravtansine (BT062)targeting multiple myeloma, anetumab ravtansine (BAY94-9343) targetingmesothelin (to treat mesothelioma), and coltuximab ravtansine (SAR3419)targeting CD19 to treat acute lymphoblastic leukemia (ALL). DM4 attachedwith the sSPDB linker is called “soravtansine” (e.g., mirvetuximabsoravtansine). Compared to DM1, the DM4 metabolites can also cross thecellular membrane and, upon diffusion, induce the extermination ofsurrounding cells, a phenomenon known as a bystander effect. Thesecytotoxic agents (e.g., DM1 and DM4) are described, e.g., in U.S. Pat.No. 8,557,966, and Bouchard, Hervé, Christian Viskov, and CarlosGarcia-Echeverria. “Antibody—drug conjugates—a new wave of cancerdrugs.” Bioorganic & medicinal chemistry letters 24.23, 5357-5363(2014), each of which is incorporated by reference in its entirety.

The disulfide linkers can display greater stability in storage and inserum than other linkers. Maytansinoids, and in particular DM1 and DM4,are cytotoxic agents that effects cell killing by preventing theformation of microtubules and depolymerization of extant microtubules.They are 100- to 1000-fold more cytotoxic than anticancer agents such asdoxorubicin, methotrexate, and vinca alkyloid, which are currently inclinical use. Alternatively, the antigen binding agents can be coupledto a taxane, a calicheamicin, a proteosome inhibitor, or a topoisomeraseinhibitor.[(1R)-3-methyl-1-[[(2S)-1-oxo-3-phenyl-2-[(3-mercaptoacetyl)amino]propyl]amino]butyl]boronic acid is a suitable proteosome inhibitor.N,N′-bis[2-(9-methylphenazine-1-carboxamido)ethyl]-1,2-ethanediamine isa suitable topoisomerase inhibitor.

In some implementations, the therapeutic agents are cytotoxins, forexample, dolastatins and auristatins, amanitins such as alpha-amanitin,beta-amanitin, gamma-amanitin or epsilon-amanitin, DNA minor groovebinding agents such as duocarmycin derivatives and modifiedpyrrolobenzodiazepine dimers, splicing inhibitors such as mayamycinanalogs or derivatives (e.g., FR901464 as set forth in U.S. Pat. No.7,825,267), tubular binding agents such as epothilone analogs andpaclitaxel and DNA damaging agents such as calicheamicins andesperamicins.

Auristatins include auristatin E, auristatin EB (AEB), auristatin EHP(AEFP), monomethyl auristatin E (MMAE). MMAE is a potent antimitoticagent that inhibits cell division by blocking the polymerization oftubulin. MMAE is 100-1000 times more potent than doxonihicin (AdriamycinRubex) and it is usually linked to a monoclonal antibody that recognizesa specific marker expression in cancer cells and directs MMAE to aspecific, targeted cancer cell. Auristatins, including MMAE, aredescribed in U.S. Patent Publication No. 20060074008; U.S. PatentPublication No. 2006022925; U.S. Pat. Nos. 7,691,962; 5,635,483; Int. J.Oncol. 15:367-72 (1999); Molecular Cancer Therapeutics, vol. 3. No. 8,pp. 921-932 (2004); Doronina, et al. “Development of potent monoclonalantibody auristatin conjugates for cancer therapy,” Nature biotechnology21.7 (2003): 778-784; and Francisco, et al, “cAC10-vcMMAE, ananti-CD30-monomethyl auristatin F conjugate with potent and selectiveantitumor activity,” Blood 102.4 (2003): 1458-1465; each of which isincorporated by reference in its entirety.

In some embodiments, the therapeutic agent is a DNA allkylator, e.g.,indolinobenzodiazepine pseudodimers (also known as IGN). The IGN familyof DNA-acting payload agents, including DGN462, are designed toeffectively alkylate DNA, while avoiding the delayed toxicity that candevelop with agents that cross-link DNA in addition to alkylating it. Adescription of IGN can be found in, e.g., Miller et al. “A New Class ofAntibody-Drug Conjugates with Potent DNA Alkylating Activity,” Molecularcancer therapeutics, molcanther-0184 (2016); U.S. Patent Publication No.20160095938; and in U.S. Pat. No. 8,765,740; each of which isincorporated by reference in its entirety.

In some embodiments, the therapeutic agents include, but are not limitedto, antimetabolites methotrexate, 6-mercaptopurine, 6-thioguanine,cytarabine, and 5-fluorouracil decarbazine), alkylating agents (e.g.,mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BCNU) andlomustine (CCNU), busulfan, dibromomannitol, streptozotocin, and cisdichlorodiamine platinum (11) (DDP) cisplatin), anthracyclines (e.g.,daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g.,dactinomycin (formerly actinomycin), bleomycin, and anthramycin (AMC)),and anti-mitotic agents (e.g., vincristine and vinblastine).

Examples for cytotoxic drugs are, without limitation, maytansinoids(e.g., emtansine, mertansine), calicheamicins (e.g., ozogamicin),auristatins (e.g., monomethyl auristatin E), pyrrolobenzodiazepines,ansamitocins, doxorubicins, daunorubicins, taxanes, bromodeoxyuridine(BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine,fluorodeoxyuridine (FudR), hydroxyurea, and cisplatin. In oneembodiment, the agent is pyrrolobenzodiazepine (PBD).

In some implementations, the therapeutic agents are enzymatically activetoxins. Enzymatically active toxins and fragments thereof include, butare not limited to, diphtheria toxin A fragment, nonbinding activefragments of diphtheria toxin, exotoxin A (from Pseudomonas aeruginosa),ricin A chain, abrin A chain, modeccin A chain, α-sacrin, certainAleurites fordii proteins, certain Dianthin proteins, Phytolaccaamericana proteins (PAP, PAPII and PAP S), Momordica charantiainhibitor, curcin, crotin, Saponaria officinalis inhibitor, gelonin,mitogillin, restrictocin, phenomycin, and enomycin. In some embodiments,the antigen binding agent is conjugated to CC-1065 (see U.S. Pat. Nos.5,475,092, 5,585,499, and 5,846,545). Procedures for preparingenzymatically active polypeptides of the immunotoxins are described,e.g., in WO 1984/03508 and WO 1985/03508, which are hereby incorporatedherein by reference in their entireties. Examples of cytotoxic moietiesthat can be conjugated to the antibodies include adriamycin,chlorambucil, daunomycin, methotrexate, neocarzinostatin, and platinum.

A therapeutic agent attached to an antibody as described herein can alsoinclude agents that are derived from, or that beneficially modulate,host biological processes, such as interferons, tumor growth factors,tumor necrosis factors, growth factors such as granulocyte macrophagecolony-stimulating factor (GM-CSF), granulocyte-colony stimulatingfactor (G-CSF), and interleukins, for example, interleukin-2,interleukin-6, interleukin-7 and interleukin-12, and the like. Atherapeutic agent attached to an antigen binding agent as describedherein may also comprise an agent that damages DNA and/or prevents cellsfrom multiplying, such as genotoxins. Genotoxins include, but are notlimited to, alkylating agents, antimetabolites, DNA cutters, DNAbinders, topoisomerase poisons, and spindle poisons. Examples ofalkylating agents include lomustine, carmustine, streptozocin,mechlorethamine, melphalan, uracil nitrogen mustard, chlorambucil,cyclosphamide, iphosphamide, cisplatin, carboplatin, mitomycin,thiotepa, dacarbazin, procarbazine, hexamethyl melamine, triethylenemelamine, busulfan, pipobroman, mitotane, and other platine derivatives.

Additional examples of cytotoxic peptides or proteins includeIdarubicin; CRM9 (e.g., FN18-CRM9, Knechtle et al., Transplantation,1997; 63:1-6); or pokeweed antiviral protein. In some embodiments, thecytotoxic protein is a bacterial toxin, e.g., diphtheria toxin (DT) orportions or variants thereof, e.g., Metl-Thr387, e.g., as described inAullo et al., EMBO J., 11(2):575-83 (1992); Abi-Habib et al., Blood,104(7):2143-2148 (2004); Perentesis et al., Proc. Nati. Acad. Sci. USA85:8386-8390 (1988); Zettlemeissl et al., Gene, 41(1):103-111 (1986); US2009/0010966; US2009/0041797; U.S. Pat. Nos. 5,843,711; 7,585,942;7,696,338; or US2008/0166375; monomethyl auristatin E; or Pseudomonasexotoxin (PE), or portions or variants thereof, e.g., as described inU.S. Pat. Nos. 4,545,985; 4,892,827; 5,458,878; 7,314,632; Song et al.,Protein Expression and Purification, 44(1):52-57 (2005); Theuer et al.,J. Biol. Chem., 267(24):16872-16877 (1992); Heimbrook et al., Proc NatlAcad Sci USA, 87(12):4697-4701 (1990); Dcbinski et al., Mol Cell Biol.,11(3):1751-1753 (1991); and Chaudhary et al., Proc. Nadl. Acad. Sci.USA, 87:308-312 (1990).

In some implementations, the cytotoxic protein is a plant toxin, e.g., aplant holotoxin (e.g., class II ribosome-inactivating proteins such asricin (e.g., deglycosylated ricin A chain (dgA)), abrin, mistletoelectin, or modeccin) or hemitoxin (class I ribosome-inactivatingproteins, e.g., PAP, saporin, bryodin 1, bouganin, or gelonin), orfragments or variants thereof that retain cytotoxic activity. See, e.g.,Neville et al., J Contr Rel., 1993; 24:133-141; Vallera, Blood, 1994;83:309-317; Vitetta et al., Immunology Today, 1993; 14:252-259; Kreitmanet al., AAPS J., 2006; 8(3):E532-E551).

In some embodiments, the cytotoxic or cytostatic agent is maytansinoid,DGN462, benzodiazepine, taxoid, CC-1065, duocarmycin, calicheamicin,dolastatin, auristatin, tomaymycin, or leptomycin. In some embodiments,the cytotoxic or cytostatic agent is mertansine/emtansine (DM1),ravtansine/soravtansine (DM4), indolino-benzodiazepine, or monomethylauristatin E (MMAE), SN-38, monomethyl auristatin phenylalanine (MMAF),doxorubicin, tubulysin (AZ13599185), pyrrolobenzodiazepine (PBD),Amberstatin-269, or topoisomerase inhibitor (DXd). Some of these agentsare described, e.g., in U.S. Pat. No. 8,557,966.

The antibody drug conjugates include an antibody or antigen-bindingfragment thereof that binds to CDCP1. The antibody can be any of theantibodies described herein. In some embodiments, the therapeutic agentin the antibody drug conjugates can be a microtubule inhibitor, e.g.,N^(2′)-deacetyl-N^(2′)-(4-mercapto-4-methyl-1-oxopentyl) maytansine(DM4), monomethyl auristatin E (MMAE), or a DNA alkylator, e.g.,indolinobenzodiazepine pseudodimers (IGN) (Table 1). The linker thatlinks the therapeutic agent and the antibody or antigen-binding fragmentcan be D-Ala-L-Ala dipeptide anilino (D-Ala-L-Ala dpa), N-succinimidyl4-(2-pyridyldithio)-2-sulfobutanoate (sSPDB), ormaleimido-caproyl-valine-citrulline (MC-VC). In some embodiments, thetherapeutic agent is DM4, and the linker is selected from the groupconsisting of D-Ala-L-Ala dpa, sSPDB, and MC-VC. In some embodiments,the therapeutic agent is MMAE, and the linker is selected from the groupconsisting of D-Ala-L-Ala dpa, sSPDB, and MC-VC. In some embodiments,the therapeutic agent is IGN, and the linker is selected from the groupconsisting of D-Ala-L-Ala dpa, sSPDB, and MC-VC.

The antibody and the antibody drug conjugates described herein can beused to treat various cancers, including, but are not limited to,cancers of the stomach, colon, rectum, mouth/pharynx, larynx, liver,pancreas, lung, breast, cervix uteri, corpus uteri, ovary, prostate,testis, bladder, skin, bone, kidney, head, neck, and throat, Hodgkinsdisease, non-Hodgkins leukemia, sarcomas, choriocarcinoma, lymphoma,brain/central nervous system, and neuroblastoma (e.g., pediatricneuroblastoma), etc. In some embodiments, the antibody drug conjugatesdescribed in the present disclosure can be used to treat breast cancer(e.g., TNBC), colon cancer, and small cell lung cancer (Table 1). Forexample, a CDCP1 antibody that is linked to DM4 by a D-Ala-L-Ala dpalinker or sSPDB linker can be used to treat triple negative breastcancer or colon cancer. In some embodiments, the CDCP1 antibodydisclosed in the present disclosure can also be linked to IGN by thelinker D-Ala-L-Ala dpa. These ADCs can be used to treat colon cancer andsmall cell lung cancer. Furthermore, the CDCP1 antibody that is linkedto MMAE by MC-VC can be used to treat TNBC and small cell lung cancer.

TABLE 1 Indication Payload Class Payload Linker Triple negativeMicrotubule DM4 Cleavable peptide (D-Ala-L- breast cancer inhibitors Aladpa) (TNBC) Charged hindered disulfide (sSPDB) MMAEmaleimido-caproyl-valine- citrulline (MC-VC) Colon Cancer DNA alkylatorIGN Cleavable peptide (D-Ala-L- Ala dpa) Microtubule DM4 Cleavablepeptide (D-Ala-L- inhibitors Ala dpa) Charged hindered disulfide (sSPDB)MMAE maleimido-caproyl-valine- citrulline (MC-VC) Small Cell DNAalkylator IGN Cleavable peptide (D-Ala-L- Lung Cancer Ala dpa) (SCLC)

The antibodies can also be coupled to high energy radiation emitters,for example, a radioisotope, such as ¹³¹I, a γ-emitter, which, whenlocalized at the target site, e.g., tumor site, results in a killing ofseveral cell diameters. See, e.g., Order, “Analysis, Results, and FutureProspective of the Therapeutic Use of Radiolabeled Antibody in CancerTherapy,” in Monoclonal Antibodies for Cancer Detection and Therapy, R.W. Baldwin et al. (eds.), pp 303-316 (Academic Press 1985). Othersuitable radioisotopes include α-emitters, such as ²¹²Bi, ²¹³Bi, and²¹¹At and β-emitters, such as ¹⁸⁶Re and ⁹⁰Y. Lu¹¹⁷ may also be used asboth an imaging and cytotoxic agent.

The antibodies can also be conjugated or fused to viral surface proteinspresent on viral particles. For example, an antigen binding agent couldbe fused (e.g., to form a fusion protein) to a viral surface protein.Alternatively, an antigen binding agent could be chemically conjugated(e.g., via a chemical linker) to a viral surface protein. Preferably,the virus is one that fuses with endocytic membranes, e.g., an influenzavirus, such that the virus is internalized along with the antigenbinding agents and thereby enters and kills the tumor cells. The viruscan be genetically engineered as a cellular toxin. For example, thevirus could express or induce the expression of genes that are toxic tocells, e.g., cell death promoting genes. Preferably, such viruses wouldbe incapable of viral replication.

In some implementations, antibodies can be conjugated with a prodrugthat is activated only when in close proximity with a prodrug activator.The prodrug activator is conjugated with a second antigen binding agent,preferably one that binds to a non-competing site on the same targetreceptor or target cell. Drug-prodrug pairs suitable for use are knownin the art, see, e.g., in Blakely et al., Cancer Research, 56:3287 3292(1996).

Additional information regarding cytotoxic agents, linkers, andproduction of antibody-drug conjugates may be found, e.g., in WO2013/055990, WO 2013/055993, WO 2012/123423, WO 2012/041805, WO2011/130613, WO 2011/130616, WO 2009/117531, WO 2007/103288, WO2007/011968, WO 2007/008603, WO 2007/008848, WO 2006/132670, WO2006/065533, WO 2005/084390, WO 2005/082023, WO 2005/081711, WO2005/077090, WO 2005/070457, WO 2004/010957, WO 2003/026577, WO2012/177837, WO 2012/1445112, WO 2012/138749, WO 2012/135517, WO2012/135522, WO 2012/128868, WO 2012/112708, WO 2012/112687, WO2012/078868, WO 2012/061590, WO 2010/141566, WO 2010/126551, WO2010/126552, WO 2010/091150, WO 2009/134870, WO 2009/134977, WO2009/134952, WO 2009/134976, WO 2009/080831, WO 2007/056550, WO2007/024536, WO 2006/086733, WO 2006/078809, WO 2006/078368, WO2005/037992, WO 2005/020883, WO 2004/110498, WO 2004/103272, WO2004/016801, WO 2004/013093, WO 2002/098883, WO 2001/024763, U.S. Pat.Nos. 9,061,074, 9,345,785, 9,428,585, 7,553,816, 8,288,352, 9,504,756,8,609,105, 7,745,394, 8,039,273, 8,992,932, 8,343,928, 8,987,209,8,697,688, 8,841,425, 9,090,629, 9,434,748, 9,353,127, 9,289,512,9,376,500, 9,125,896, 9,498,541, U.S. 9,469,655, U.S. Pat. Nos.9,375,488, and 9,289,509, each of which are incorporated herein byreference in its entirety.

Other agents include, but are not limited to, abrin (e.g. abrin Achain), alpha toxin, Aleurites fordii proteins, amatoxin, crotin,curcin, dianthin proteins, diptheria toxin (e.g. diphtheria A chain andnonbinding active fragments of diphtheria toxin), deoxyribonuclease(Dnase), gelonin, mitogellin, modeccin A chain, Momordica charantiainhibitor, neomycin, onconase, phenomycin, Phytolaca americana proteins(PAPI, PAPII, and PAP-S), pokeweed antiviral protein, Pseudomonasendotoxin, Pseudomonas exotoxin (e.g. exotoxin A chain (from Pseudomonasaeruginosa)), restrictocin, ricin A chain, ribonuclease (Rnase),Sapaonaria officinalis inhibitor, saporin, alpha-sarcin, Staphylcoccalenterotoxin-A, tetanus toxin, cisplatin, carboplatin, and oxaliplatin(Eloxatin, Sanofi Aventis), proteasome inhibitors (e.g. PS-341[bortezomib or Velcade]), HDAC inhibitors (vorinostat (Zolinza, Merck &Company, Inc.)), belinostat, entinostat, mocetinostat, andpanobinostat), COX-2 inhibitors, substituted ureas, heat shock proteininhibitors (e.g. Geldanamycin and its numerous analogs), adrenocorticalsuppressants, and the tricothecenes. (See, for example, WO 93/21232).Other agents also include asparaginase (Espar, Lundbeck Inc.),hydroxyurea, levamisole, mitotane (Lysodren, Bristol-Myers Squibb), andtretinoin (Renova, Valcant Pharmaceuticals Inc.).

It should be noted that the aforementioned groups of drug moieties thatmay be used in the anti-CDCP1 ADCs are not exclusive, in that certainexamples of drugs may be found in more than one category, e.g.,ansamitocins are both mitotic inhibitors and antitumor antibiotics.

All stereoisomers of the above drug moieties are contemplated for useherein, i.e. any combination of R and S configurations at the chiralcarbons of D.

The above agents (i.e., naked agents not conjugated to an antibody) mayalso be used in combination therapies with the anti-CDCP1 antibodiesdescribed herein. In one embodiment, anti-CDCP1 antibodies or ADCs areused with any of the foregoing agents in a combination therapy to treatcancer, where the agent is administered prior to, at the same time as,or following administration of the anti-CDCP1 antibody or ADC to thesubject.

B. Anti-CDCP1 ADCs: Exemplary Linkers

An anti-CDCP1 ADC comprises an anti-CDCP1 antibody and at least onedrug(s), whereby the antibody and the at least one drug are conjugatedby a linker. The term “linker,” as used herein, refers to a chemicalmoiety that may be bifunctional or multifunctional, and is used toattach an antibody to a drug moiety. A linker may include oneconjugating component or may include multiple components.

For example, the linker may include a spacer, which is a moiety thatextends the drug linkage to avoid, for example, shielding the activesite of the antibody or improving the solubility of the ADC. Otherexamples of components of linkers include a stretcher unit and an aminoacid unit.

Two methods are commonly used for conjugating drugs to antibodies:alkylation of reduced interchain cysteine disulfides through anenzymatically non-cleavable maleimido or simple and cleavable disulfidelinker, and acylation of lysines by cleavable linear amino acids.

In one aspect, a linker covalently attaches an antibody to a drugmoiety. An ADC is prepared using a linker having reactive functionalityfor binding to the antibody and the drug. For example, a cysteine thiol,or an amine, e.g., N-terminus or amino acid side chain such as lysine,of the antibody may form a bond with a functional group of the linker.

In one aspect, a linker has a functionality that is capable of reactingwith a free cysteine present on an antibody to form a covalent bond.Nonlimiting exemplary such reactive functionalities include maleimide,haloacetamides, □-haloacetyl, activated esters such as succinimideesters, 4-nitrophenyl esters, pentafluorophenyl esters,tetrafluorophenyl esters, anhydrides, acid chlorides, sulfonylchlorides, isocyanates, and isothiocyanates. See, e.g., the conjugationmethod at page 766 of Klussman, et al (2004), Bioconjugate Chemistry15(4):765-773.

In some embodiments, a linker has a functionality that is capable ofreacting with an electrophilic group present on an antibody. Exemplarysuch electrophilic groups include, but are not limited to, aldehyde andketone carbonyl groups. In some embodiments, a heteroatom of thereactive functionality of the linker can react with an electrophilicgroup on an antibody and form a covalent bond to an antibody unit.Nonlimiting exemplary such reactive functionalities include, but are notlimited to, hydrazide, oxime, amino, hydrazine, thiosemicarbazone,hydrazine carboxylate, and arylhydrazide.

Suitable linkers include, for example, cleavable and non-cleavablelinkers. A linker may be a “cleavable linker,” facilitating release of adrug. Nonlimiting exemplary cleavable linkers include acid-labilelinkers (e.g., comprising hydrazone), protease-sensitive (e.g.,peptidase-sensitive) linkers, photolabile linkers, ordisulfide-containing linkers (Chari et al., Cancer Research 52:127-131(1992); U.S. Pat. No. 5,208,020). A cleavable linker is typicallysusceptible to cleavage under intracellular conditions. Suitablecleavable linkers include, for example, a peptide linker cleavable by anintracellular protease, such as lysosomal protease or an endosomalprotease. In exemplary embodiments, the linker can be a dipeptidelinker, such as a a valine-citrulline (Val-Cit) linker, an Ala-Vallinker, or a Phe-Lys linker. These linkers are known in the art, and aredescribed, e.g., in U.S. Pat. No. 6,214,345. One advantage of usingintracellular proteolytic release of the therapeutic agent is that theagent is typically attenuated when conjugated and the serum stabilitiesof the conjugates are typically high.

Linkers are preferably stable extracellularly in a sufficient manner tobe therapeutically effective. Before transport or delivery into a cell,the ADC is preferably stable and remains intact, i.e. the antibodyremains conjugated to the drug moiety. Linkers that are stable outsidethe target cell may be cleaved at some efficacious rate once inside thecell. Thus, an effective linker will: (i) maintain the specific bindingproperties of the antibody; (ii) allow delivery, e.g., intracellulardelivery, of the drug moiety; and (iii) maintain the therapeutic effect,e.g., cytotoxic effect, of a drug moiety.

In one embodiment, the linker is cleavable under intracellularconditions, such that cleavage of the linker sufficiently releases thedrug from the antibody in the intracellular environment to betherapeutically effective. In some embodiments, the cleavable linker ispH-sensitive, i.e., sensitive to hydrolysis at certain pH values.Typically, the pH-sensitive linker is hydrolyzable under acidicconditions. For example, an acid-labile linker that is hydrolyzable inthe lysosome (e.g., a hydrazone, semicarbazone, thiosemicarbazone,cis-aconitic amide, orthoester, acetal, ketal, or the like) can be used.(See, e.g., U.S. Pat. Nos. 5,122,368; 5,824,805; 5,622,929; Dubowchikand Walker, 1999, Pharm. Therapeutics 83:67-123; Neville et al., 1989,Biol. Chem. 264:14653-14661.) Such linkers are relatively stable underneutral pH conditions, such as those in the blood, but are unstable atbelow pH 5.5 or 5.0, the approximate pH of the lysosome. In certainembodiments, the hydrolyzable linker is a thioether linker (such as,e.g., a thioether attached to the therapeutic agent via an acylhydrazonebond (see, e.g., U.S. Pat. No. 5,622,929).

In other embodiments, the linker is cleavable under reducing conditions(e.g., a disulfide linker). A variety of disulfide linkers are known inthe art, including, for example, those that can be formed using SATA(N-succinimidyl-5-acetylthioacetate), SPDP(N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB(N-succinimidyl-3-(2-pyridyldithio)butyrate) and SMPT(N-succinimidyloxycarbonyl-alpha-methyl-alpha-(2-pyridyl-dithio)toluene),SPDB and SMPT. (See, e.g., Thorpe et al., 1987, Cancer Res.47:5924-5931; Wawrzynczak et al., In Immunoconjugates: AntibodyConjugates in Radioimagery and Therapy of Cancer (C. W. Vogel ed.,Oxford U. Press, 1987. See also U.S. Pat. No. 4,880,935.).

In some embodiments, the linker is cleavable by a cleaving agent, e.g.,an enzyme, that is present in the intracellular environment (e.g.,within a lysosome or endosome or caveolea). The linker can be, e.g., apeptidyl linker that is cleaved by an intracellular peptidase orprotease enzyme, including, but not limited to, a lysosomal or endosomalprotease. In some embodiments, the peptidyl linker is at least two aminoacids long or at least three amino acids long. Cleaving agents caninclude cathepsins B and D and plasmin, all of which are known tohydrolyze dipeptide drug derivatives resulting in the release of activedrug inside target cells (see, e.g., Dubowchik and Walker, 1999, Pharm.Therapeutics 83:67-123). Most typical are peptidyl linkers that arecleavable by enzymes that are present in CDCP1-expressing cells.Examples of such linkers are described, e.g., in U.S. Pat. No.6,214,345, incorporated herein by reference in its entirety and for allpurposes. In a specific embodiment, the peptidyl linker cleavable by anintracellular protease is a Val-Cit linker or a Phe-Lys linker (see,e.g., U.S. Pat. No. 6,214,345, which describes the synthesis ofdoxorubicin with the val-cit linker). One advantage of usingintracellular proteolytic release of the therapeutic agent is that theagent is typically attenuated when conjugated and the serum stabilitiesof the conjugates are typically high.

In other embodiments, the linker is a malonate linker (Johnson et al.,1995, Anticancer Res. 15:1387-93), a maleimidobenzoyl linker (Lau etal., 1995, Bioorg-Med-Chem. 3(10):1299-1304), or a 3′-N-amide analog(Lau et al., 1995. Bioorg-Med-Chem. 3(10): 1305-12).

In some implementations, the antigen binding agents can be directlyconjugated to radioisotopes or may comprise macrocyclic chelators usefulfor conjugating radio-metal ions. In some embodiments, the macrocyclicchelator is 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″-tetraacetic acid(DOTA), which can be attached to the antibody via a linker molecule.These linkers are known in the art and are described, e.g., in Denardoet al., 1998, Clin Cancer Res., 4:2483; Peterson et al., 1999,Bioconjug. Chem., 10:553; and Zimmerman et al., 1999, Nucl. Med. Biol.,26:943, each of which is incorporated by reference in its entirety.

In yet other embodiments, the linker unit is not cleavable and the drugis released, for example, by antibody degradation. See U.S. PublicationNo. 20050238649 incorporated by reference herein in its entirety. An ADCcomprising a non-cleavable linker may be designed such that the ADCremains substantially outside the cell and interacts with certainreceptors on a target cell surface such that the binding of the ADCinitiates (or prevents) a particular cellular signaling pathway.

In some embodiments, the linker is N-succinimidyl4-(2-pyridyldithio)pentanoate (SPP), N-succinimidyl4-(2-pyridyldithio)-2-sulfopentanoate (sulfo-SPP), N-succinimidyl4-(maleimidomethyl) cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl4-(maleimidomethyl) cyclohexanecarboxylate (sulfoSMCC),N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), orN-succinimidyl-[(N-maleimidopropionamido)-tetraethyleneglycol]ester(NHS-PEG4-maleimide). Some of these linkers are described, e.g., in U.S.Pat. No. 8,557,966.

In some embodiments, the linker is substantially hydrophilic linker(e.g., PEG4Mal and sulfo-SPDB). A hydrophilic linker may be used toreduce the extent to which the drug may be pumped out of resistantcancer cells through MDR (multiple drug resistance) or functionallysimilar transporters.

In other embodiments, upon cleavage, the linker functions to directly orindirectly inhibit cell growth and/or cell proliferation. For example,in some embodiments, the linker, upon cleavage, can function as anintercalating agent, thereby inhibiting macromolecular biosynthesis(e.g. DNA replication, RNA transcription, and/or protein synthesis).

In other embodiments, the linker is designed to facilitate bystanderkilling (the killing of neighboring cells) through diffusion of thelinker-drug and/or the drug alone to neighboring cells. In other,embodiments, the linker promotes cellular internalization.

The presence of a sterically hindered disulfide can increase thestability of a particular disulfide bond, enhancing the potency of theADC. Thus, in one embodiment, the linker includes a sterically hindereddisulfide linkage. A sterically hindered disulfide refers to a disulfidebond present within a particular molecular environment, wherein theenvironment is characterized by a particular spatial arrangement ororientation of atoms, typically within the same molecule or compound,which prevents or at least partially inhibits the reduction of thedisulfide bond. Thus, the presence of bulky (or sterically hindering)chemical moieties and/or bulky amino acid side chains proximal to thedisulfide bond prevents or at least partially inhibits the disulfidebond from potential interactions that would result in the reduction ofthe disulfide bond.

Notably, the aforementioned linker types are not mutually exclusive. Forexample, in one embodiment, the linker used in the anti-CDCP1 ADCsdescribed herein is a non-cleavable linker that promotes cellularinternalization.

In some embodiments, the ADC has the following formula (formula I):Ab-(L-D)_(n)  (I)or a pharmaceutically acceptable salt or solvate thereof; wherein Ab isthe antibody, e.g., anti-CDCP1 antibody, and (L-D) is a Linker-Drugmoiety. The Linker-Drug moiety is made of L- which is a Linker, and -D,which is a drug moiety having, for example, cytostatic, cytotoxic, orotherwise therapeutic activity against a target cell, e.g., a cellexpressing CDCP1; and n is an integer from 1 to 20.

In some embodiments, n ranges from 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to4, 1 to 3, 1 to 2, or is I.

In some embodiments, the -D moieties are the same. In yet anotherembodiment, the -D moieties are different.

In some embodiments, a linker component comprises an “amino acid unit.”In some such embodiments, the amino acid unit allows for cleavage of thelinker by a protease, thereby facilitating release of the drug from theimmunoconjugate upon exposure to intracellular proteases, such aslysosomal enzymes (Doronina et al. (2003) Nat. Biotechnol. 21:778-784).Exemplary amino acid units include, but are not limited to, dipeptides,tripeptides, tetrapeptides, and pentapeptides. Exemplary dipeptidesinclude, but are not limited to, valine-citrulline (vc or val-cit),alanine-phenylalanine (af or ala-phe); phenylalanine-lysine (fk orphe-lys); phenylalanine-homolysine (phe-homolys); andN-methyl-valine-citrulline (Me-val-cit). Exemplary tripeptides include,but are not limited to, glycine-valine-citrulline (gly-val-cit) andglycine-glycine-glycine (gly-gly-gly). An amino acid unit may compriseamino acid residues that occur naturally and/or minor amino acids and/ornon-naturally occurring amino acid analogs, such as citrulline Aminoacid units can be designed and optimized for enzymatic cleavage by aparticular enzyme, for example, a tumor-associated protease, cathepsinB, C and D, or a plasmin protease.

In one embodiment, the amino acid unit is valine-citrulline (vc orval-cit). In another aspect, the amino acid unit is phenylalanine-lysine(i.e., fk). In yet another aspect of the amino acid unit, the amino acidunit is N-methylvaline-citrulline. In yet another aspect, the amino acidunit is 5-aminovaleric acid, homo phenylalanine lysine,tetraisoquinolinecarboxylate lysine, cyclohexylalanine lysine,isonepecotic acid lysine, beta-alanine lysine, glycine serine valineglutamine and isonepecotic acid.

Another approach for the generation of ADCs involves the use ofheterobifunctional cross-linkers which link the anti-CDCP1 antibody tothe drug moiety. Examples of cross-linkers that may be used includeN-succinimidyl 4-(5-nitro-2-pyridyldithio)-pentanoate or the highlywater-soluble analog N-sulfosuccinimidyl4-(5-nitro-2-pyridyldithio)-pentanoate,N-succinimidyl-4-(2-pyridyldithio) butyrate (SPDB),N-succinimidyl-4-(5-nitro-2-pyridyldithio) butyrate (SNPB), andN-sulfosuccinimidyl-4-(5-nitro-2-pyridyldithio) butyrate (SSNPB),N-succinimidyl-4-methyl-4-(5-nitro-2-pyridyldithio)pentanoate (SMNP),N-succinimidyl-4-(5-N,N-dimethylcarboxamido-2-pyridyldithio) butyrate(SCPB) or N-sulfosuccinimidyl4-(5-N,N-dimethylcarboxamido-2-pyridyldithio) butyrate (SSCPB)). Theantibodies may be modified with the cross-linkers N-succinimidyl4-(5-nitro-2-pyridyldithio)-pcntanoate, N-sulfosuccinimidyl4-(5-nitro-2-pyridyldithio)-pentanoate, SPDB, SNPB, SSNPB, SMNP, SCPB,or SSCPB can then react with a small excess of a particular drug thatcontains a thiol moiety to give excellent yields of an ADC (see alsoU.S. Pat. No. 6,913,748, incorporated by reference herein).

In one embodiment, charged linkers (also referred to as pro-chargedlinkers) are used to conjugate anti-CDCP1 antibodies to drugs to formADCs. Charged linkers include linkers that become charged after cellprocessing. The presence of a charged group(s) in the linker of aparticular ADC or on the drug after cellular processing provides severaladvantages, such as (i) greater water solubility of the ADC, (ii)ability to operate at a higher concentration in aqueous solutions, (iii)ability to link a greater number of drug molecules per antibody,potentially resulting in higher potency, (iv) potential for the chargedconjugate species to be retained inside the target cell, resulting inhigher potency, and (v) improved sensitivity of multidrug resistantcells, which would be unable to export the charged drug species from thecell. Examples of some suitable charged or pro-charged cross-linkers andtheir synthesis are shown in FIGS. 1 to 10 of U.S. Pat. No. 8,236,319,and are incorporated by reference herein. Preferably, the charged orpro-charged cross-linkers are those containing sulfonate, phosphate,carboxyl or quaternary amine substituents that significantly increasethe solubility of the ADCs, especially for ADCs with 2 to 20 conjugateddrugs. Conjugates prepared from linkers containing a pro-charged moietywould produce one or more charged moieties after the conjugate ismetabolized in a cell.

Additional examples of linkers that can be used with the compositionsand methods include valine-citrulline; maleimidocaproyl; amino benzoicacids; p-aminobenzylcarbamoyl (PAB); lysosomal enzyme-cleavable linkers;maleimidocaproyl-polyethylene glycol (MC(PEG) 6-OH); N-methyl-valinecitrulline; N-succinimidyl4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC); N-Succinimidyl4-(2-pyridyldithio)butanoate (SPDB); and N-Succinimidyl4-(2-pyridylthio)pentanoate (SPP) (See also US 2011/0076232). Anotherlinker for use includes an avidin-biotin linkage to provide anavidin-biotin-containing ADC (See also U.S. Pat. No. 4,676,980, PCTpublication Nos. WO1992/022332A2, WO1994/016729A1, WO1995/015770A1,WO1997/031655A2, WO1998/035704A1, WO1999/019500A1, WO2001/09785A2,WO2001/090198A1, WO2003/093793A2, WO2004/050016A2, WO2005/081898A2,WO2006/083562A2, WO2006/089668A1, WO2007/150020A1, WO2008/135237A1,WO2010/111198A1, WO2011/057216A1, WO2011/058321A1, WO2012/027494A1, andEP77671B1), wherein some such linkers are resistant to biotinidasecleavage. Additional linkers that may be used include a cohesin/dockerinpair to provide a cohesion-dockerin-containing ADC (See PCT publicationNos. WO2008/097866A2, WO2008/097870A2, WO2008/103947A2, andWO2008/103953A2).

Additional linkers may contain non-peptide polymers (examples include,but are not limited to, polyethylene glycol, polypropylene glycol,polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, dextran,polyvinyl ethyl ether, PLA (poly(lactic acid)), PLGA (poly(lacticacid-glycolic acid)), and combinations thereof, wherein a preferredpolymer is polyethylene glycol) (See also PCT publication No.WO2011/000370). Additional linkers are also described in WO 2004-010957,U.S. Publication No. 20060074008, U.S. Publication No. 20050238649, andU.S. Publication No. 20060024317, each of which is incorporated byreference herein in its entirety).

For an ADC comprising a maytansinoid, many positions on maytansinoidscan serve as the position to chemically link the linking moiety. In oneembodiment, maytansinoids comprise a linking moiety that contains areactive chemical group are C-3 esters of maytansinol and its analogswhere the linking moiety contains a disulfide bond and the chemicalreactive group comprises a N-succinimidyl or N-sulfosuccinimidyl ester.For example, the C-3 position having a hydroxyl group, the C-14 positionmodified with hydroxymethyl, the C-15 position modified with hydroxy andthe C-20 position having a hydroxy group are all useful. The linkingmoiety most preferably is linked to the C-3 position of maytansinol.

The conjugation of the drug to the antibody via a linker can beaccomplished by any technique known in the art. A number of differentreactions are available for covalent attachment of drugs and linkers toantibodies. This may be accomplished by reaction of the amino acidresidues of the antibody, including the amine groups of lysine, the freecarboxylic acid groups of glutamic and aspartic acid, the sulfhydrylgroups of cysteine and the various moieties of the aromatic amino acids.One of the most commonly used non-specific methods of covalentattachment is the carbodiimide reaction to link a carboxy (or amino)group of a compound to amino (or carboxy) groups of the antibody.Additionally, bifunctional agents such as dialdehydes or imidoestershave been used to link the amino group of a compound to amino groups ofan antibody. Also available for attachment of drugs to antibodies is theSchiff base reaction. This method involves the periodate oxidation of adrug that contains glycol or hydroxy groups, thus forming an aldehydewhich is then reacted with the binding agent. Attachment occurs viaformation of a Schiff base with amino groups of the antibody.Isothiocyanates can also be used as coupling agents for covalentlyattaching drugs to antibodies. Other techniques are known to the skilledartisan and within the scope of the disclosure.

In certain embodiments, an intermediate, which is the precursor of thelinker, is reacted with the drug under appropriate conditions. Incertain embodiments, reactive groups are used on the drug or theintermediate. The product of the reaction between the drug and theintermediate, or the derivatized drug, is subsequently reacted with theanti-CDCP1 antibody under appropriate conditions. The synthesis andstructure of exemplary linkers, stretcher units, amino acid units,self-immolative spacer units are described in U.S. Patent ApplicationPublication Nos. 20030083263, 20050238649 and 20050009751, each if whichis incorporated herein by reference.

Stability of the ADC may be measured by standard analytical techniquessuch as mass spectroscopy, HPLC, and the separation/analysis techniqueLC/MS.

Additional information regarding cytotoxic agents, linkers, andproduction of antibody-drug conjugates may be found, e.g., in WO2013/055990, WO 2013/055993, WO 2012/123423, WO 2012/041805, WO2011/130613, WO 2011/130616, WO 2009/117531, WO 2007/103288, WO2007/011968, WO 2007/008603, WO 2007/008848, WO 2006/132670, WO2006/065533, WO 2005/084390, WO 2005/082023, WO 2005/081711, WO2005/077090, WO 2005/070457, WO 2004/010957, WO 2003/026577, WO2012/177837, WO 2012/1445112, WO 2012/138749, WO 2012/135517, WO2012/135522, WO 2012/128868, WO 2012/112708, WO 2012/112687, WO2012/078868, WO 2012/061590, WO 2010/141566, WO 2010/126551, WO2010/126552, WO 2010/091150, WO 2009/134870, WO 2009/134977, WO2009/134952, WO 2009/134976, WO 2009/080831, WO 2007/056550, WO2007/024536, WO 2006/086733, WO 2006/078809, WO 2006/078368, WO2005/037992, WO 2005/020883, WO 2004/110498, WO 2004/103272, WO2004/016801, WO 2004/013093, WO 2002/098883, WO 2001/024763, U.S. Pat.Nos. 9,061,074, 9,345,785, 9,428,585, 7,553,816, 8,288,352, 9,504,756,8,609,105, 7,745,394, 8,039,273, 8,992,932, 8,343,928, 8,987,209,8,697,688, 8,841,425, 9,090,629, 9,434,748, 9,353,127, 9,289,512,9,376,500, 9,125,896, 9,498,541, 9,469,655, 9,375,488, and 9,289,509,the entire contents of each of which are incorporated herein byreference.

IV. Uses of Anti-CDCP1 Antibodies and Anti-CDCP1 ADCs

The antibodies and antibody portions (and ADCs) preferably are capableof neutralizing human CDCP1 activity both in vivo and in vitro.Accordingly, such antibodies and antibody portions can be used toinhibit hCDCP1 activity, e.g., in a cell culture containing hCDCP1, inhuman subjects or in other mammalian subjects having CDCP1 with which anantibody disclosed herein cross-reacts. In one embodiment, thedisclosure provides a method for inhibiting hCDCP1 activity comprisingcontacting hCDCP1 with an antibody or antibody portion such that hCDCP1activity is inhibited. For example, in a cell culture containing, orsuspected of containing hCDCP1, an antibody or antibody portion can beadded to the culture medium to inhibit hCDCP1 activity in the culture.

In another embodiment, disclosed herein is a method for reducing hCDCP1activity in a subject, advantageously from a subject suffering from aCDCP1 associated disorder, e.g., cancer such as breast cancer, lungcancer, small cell lung cancer, liver cancer, pancreatic cancer, ovariancancer, kidney cancer, and colon cancer, or a disorder in which CDCP1activity is detrimental. The disclosure provides methods for reducingCDCP1 activity in a subject suffering from such a disease or disorder,which method comprises administering to the subject an antibody orantibody portion of the disclosure such that CDCP1 activity in thesubject is reduced. Preferably, the CDCP1 is human CDCP1, and thesubject is a human subject. Alternatively, the subject can be a mammalexpressing a CDCP1 to which antibodies of the disclosure are capable ofbinding. Still further the subject can be a mammal into which CDCP1 hasbeen introduced (e.g., by administration of CDCP1 or by expression of aCDCP1 transgene). Antibodies of the disclosure can be administered to ahuman subject for therapeutic purposes. Moreover, antibodies of thedisclosure can be administered to a non-human mammal expressing a CDCP1with which the antibody is capable of binding for veterinary purposes oras an animal model of human disease. Regarding the latter, such animalmodels may be useful for evaluating the therapeutic efficacy ofantibodies of the disclosure (e.g., testing of dosages and time coursesof administration).

As used herein, the term “a disorder in which CDCP1 activity isdetrimental” is intended to include diseases and other disorders inwhich the presence of CDCP1 in a subject suffering from the disorder hasbeen shown to be or is suspected of being either responsible for thepathophysiology of the disorder or a factor that contributes to aworsening of the disorder. Accordingly, a disorder in which CDCP1activity is detrimental is a disorder in which reduction of CDCP1activity is expected to alleviate the symptoms and/or progression of thedisorder. Such disorders may be evidenced, for example, by an increasein the concentration of CDCP1 in a biological cell, fluid or tissue of asubject suffering from the disorder (e.g., an increase in theconcentration of CDCP1 in a tumor, scrum, plasma, synovial fluid, etc.of the subject), which can be detected, for example, using an anti-CDCP1antibody as described above.

Non-limiting examples of disorders that can be treated with theantibodies, or antigen binding fragments thereof, include thosedisorders discussed below. For example, suitable disorders include, butare not limited to, a variety of cancers including, but not limited to,breast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer.

In one aspect, the disclosure features methods that includeadministering an antibody or composition (e.g., a cell composition,antibody-drug conjugate, or antibody-radioisotope conjugate) disclosedherein to a subject in need thereof (e.g., a subject having, oridentified or diagnosed as having, a cancer characterized by expressionor (overexpression) of CDCP1, or a cancer characterized by the presenceof CDCP1 on the surface of the cancer cells (e.g., a subject identifiedusing any of the methods described herein), e.g., breast cancer (e.g.,triple-negative breast cancer), carcinoid cancer, cervical cancer,endometrial cancer, glioma, head and neck cancer, liver cancer, lungcancer, small cell lung cancer, lymphoma, melanoma, ovarian cancer,pancreatic cancer, prostate cancer, renal cancer, colorectal cancer,gastric cancer, testicular cancer, thyroid cancer, bladder cancer,urethral cancer, or hematologic malignancy. In some implementations, thesubject is identified as being a subject that expresses CDCP1 (e.g.,using any of the methods described herein) or has an elevated level of aCDCP1 protein (e.g., as compared to reference level, e.g., a level of aCDCP1 protein produced by a healthy subject, a level of a CDCP1 proteinproduced by a non-cancerous, e.g., primary, cell, or a threshold levelof a CDCP1 protein, in which a determined level of a CDCP1 protein thatis above this value indicates that the subject should be administered anantibody described herein, an antibody-drug conjugate described herein,and/or other therapy). In some embodiments, the compositions and methodsdisclosed herein can be used for treatment of patients at risk for acancer.

In some implementations, the subject has a cancer characterized by thepresence of CDCP1 on the cancer cell surface. In some cases, CDCP1proteins are translocated from the cytoplasm to the cell surface, or tocell surface CDCP1 rafts. Thus, the quantity of CDCP1 in a cancer cellmay be similar to the quantity of CDCP1 in a normal cell. In some cases,the tumor cells expressing CDCP1 proliferate, therefore the quantity ofCDCP1 in tumor tissue is higher relative to the quantity of CDCP1 innormal tissue.

In yet another aspect, the disclosure features methods of inhibiting ordecreasing proliferation of a cell (e.g., a cell that expresses (e.g.,overexpresses) CDCP1, or a cell characterized by the presence of CDCP1on the cell surface) that include contacting the cell with an antibody,antibody-drug conjugate, nucleic acid, composition, or cell disclosedherein. In another aspect, the disclosure features methods of inhibitingor decreasing proliferation of a cancer cell (e.g., a cancer cell thatoverexpresses CDCP1, or a cancer cell characterized by having CDCP1 onthe cell surface) that include contacting the cancer cell with anantibody, antibody-drug conjugate, nucleic acid, composition, or celldisclosed herein. A cell can be identified as overexpressing CDCP1protein using any of the methods described herein. In some embodiments,a cell can also be identified as having CDCP1 on the cell surface usingany of the methods described herein. In some implementations, the celldeath induced by the antibody is complement-dependent cytotoxicityand/or cell-dependent cell cytotoxicity. In some implementations, theantibody is conjugated to a therapeutic agent (e.g., a cytotoxic agent)and the cell death is induced by the endocytosis of the antibody intothe cancer cells, which in turn triggers the death of the cancer cells.In some implementations, the methods include the step of administeringto a subject (e.g., a subject in need thereof) an effective amount of anantibody, nucleic acid, composition, or cell disclosed herein, therebyselectively inducing cell death.

Other examples of cancers that may be treated using the compositions andmethods disclosed herein include, but are not limited to carcinoma,lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. Moreparticular examples of such cancers include but are not limited tobreast cancer (Luminal A, TNBC, Ductal), prostate cancer, squamous celltumors, squamous cell carcinoma (e.g., squamous cell lung cancer orsquamous cell head and neck cancer), neuroendocrine tumors, urothelialcancer, vulvar cancer, mesothelioma, liver cancer, bone cancer,pancreatic cancer, skin cancer, cancer of the head or neck, lung cancer,small cell lung cancer, non-small cell lung cancer, cutaneous orintraocular malignant melanoma, renal cancer, uterine cancer, ovariancancer, colorectal cancer, colon cancer, rectal cancer, cancer of theanal region, stomach cancer, testicular cancer, uterine cancer,carcinoma of the fallopian tubes, carcinoma of the endometrium,carcinoma of the cervix, carcinoma of the vagina, carcinoma of thevulva, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of thesmall intestine, cancer of the endocrine system, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, solid tumors of childhood,lymphocytic lymphoma, cancer of the bladder, cancer of the kidney orureter, carcinoma of the renal pelvis, neoplasm of the central nervoussystem (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axistumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma,epidermoid cancer, environmentally induced cancers including thoseinduced by asbestos, hematologic malignancies including, for example,multiple myeloma, B-cell lymphoma, Hodgkin lymphoma/primary mediastinalB-cell lymphoma, non-Hodgkin's lymphomas, acute myeloid lymphoma,chronic myelogenous leukemia, chronic lymphoid leukemia, follicularlymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma,immunoblastic large cell lymphoma, precursor B-Iymphoblastic lymphoma,mantle cell lymphoma, acute lymphoblastic leukemia, mycosis fungoides,anaplastic large cell lymphoma, T-cell lymphoma, and precursorT-lymphoblastic lymphoma, and any combinations of said cancers. PVNS,acute myeloid leukemia, adrenocortico carcinoma, ladder urothelialcarcinoma, cervical squamous cell carcinoma, endocervicaladenocarcinoma, diffuse large B cell lymphoma, glioblastoma multiforme,chronic lymphocytic leukemia, brain lower grade glioma, head and necksquamous cell carcinoma, hepatocellular carcinoma, lung adenocarcinoma,large squamous cell carcinoma, cutaneous melanoma, ovarial serouscystadenocarcinoma, gastric cancer, soft tissue sarcoma, testicular germcell cancer, thymoma, thyroid carcinoma, uterine corpus endometrialcarcinoma, uterine carcinosarcoma, kidney renal clear cell carcinoma,and kidney renal papillary cell carcinoma. The present invention is alsoapplicable to treatment of metastatic cancers.

In one embodiment, the antibodies and ADCs disclosed herein are used totreat a solid tumor, e.g., inhibit growth of or decrease size of a solidtumor, overexpressing CDCP1 or which is CDCP1 positive. In anotherembodiment, the antibodies and ADCs disclosed herein are used to treatbreast cancer, lung cancer, small cell lung cancer, liver cancer,pancreatic cancer, ovarian cancer, kidney cancer, and colon cancer.Diseases and disorders described herein may be treated by anti-CDCP1antibodies or ADCs, as well as pharmaceutical compositions comprisingsuch anti-CDCP1 antibodies or ADCs.

In certain embodiments, the antibodies and ADCs disclosed herein areadministered to a subject in need thereof in order to treat advancedcancers likely to exhibit elevated levels of CDCP1.

In certain embodiments, the disclosure includes a method for inhibitingor decreasing solid tumor growth in a subject having a solid tumor, saidmethod comprising administering an anti-CDCP1 antibody or ADC describedherein, to the subject having the solid tumor, such that the solid tumorgrowth is inhibited or decreased. In further embodiments, the solidtumor is an CDCP1 expressing solid tumor. In certain embodiments theanti-CDCP1 antibodies or ADCs described herein are administered to asubject having cancer, alone or in combination with an additional agent,e.g., radiation and/or chemotherapy, an immune checkpoint inhibitor, ora PARP inhibitor.

In certain embodiments, the disclosure includes a method for inhibitingor decreasing solid tumor growth in a subject having a solid tumor whichwas identified as a CDCP1 expressing or CDCP1 expressing tumor, saidmethod comprising administering an anti-CDCP1 antibody or ADC describedherein, to the subject having the solid tumor, such that the solid tumorgrowth is inhibited or decreased. Methods for identifying CDCP1expressing tumors are known in the art, and include FDA-approved testsand validation assays. For example, these assays may use primers thatare specific for the CDCP1 gene and/or cDNA and result in theamplification of the CDCP1 gene/cDNA, or a portion thereof. Theamplified PCR products may be subsequently analyzed, for example, by gelelectrophoresis using standard methods known in the art to determine thesize of the PCR products. Such tests may be used to identify tumors thatmay be treated with the methods and compositions described herein.

In another aspect, this application features a method of treating (e.g.,curing, suppressing, ameliorating, delaying or preventing the onset of,or preventing recurrence or relapse of) or preventing a CDCP1-associateddisorder, in a subject. The method includes: administering to thesubject a CDCP1 binding agent (particularly an antagonist), e.g., ananti-CDCP1 antibody or fragment thereof as described herein, in anamount sufficient to treat or prevent the CDCP1-associated disorder. TheCDCP1 antagonist, e.g., the anti-CDCP1 antibody or fragment thereof, canbe administered to the subject, alone or in combination with othertherapeutic modalities as described herein.

Antibodies or ADCs, or antigen binding portions thereof, can be usedalone or in combination to treat such diseases. It should be understoodthat the antibodies or antigen binding portion thereof can be used aloneor in combination with an additional agent, e.g., a therapeutic agent,said additional agent being selected by the skilled artisan for itsintended purpose. For example, the additional agent can be a therapeuticagent art-recognized as being useful to treat the disease or conditionbeing treated by the antibody. The additional agent also can be an agentthat imparts a beneficial attribute to the therapeutic composition,e.g., an agent which affects the viscosity of the composition.

It should further be understood that the combinations which are to beincluded within this disclosure are those combinations useful for theirintended purpose. The agents set forth below are illustrative forpurposes and not intended to be limited. The combinations, which arepart of this disclosure, can be the antibodies of the disclosure and atleast one additional agent selected from the lists below. Thecombination can also include more than one additional agent, e.g., twoor three additional agents if the combination is such that the formedcomposition can perform its intended function.

The combination therapy can include one or more CDCP1 antagonists, e.g.,anti-CDCP1 antibodies or fragments thereof, formulated with, and/orco-administered with, one or more additional therapeutic agents, e.g.,one or more cytokine and growth factor inhibitors, immunosuppressants,anti-inflammatory agents (e.g., systemic anti-inflammatory agents),anti-fibrotic agents, metabolic inhibitors, enzyme inhibitors, and/orcytotoxic or cytostatic agents, mitotic inhibitors, antitumorantibiotics, immunomodulating agents, vectors for gene therapy,alkylating agents, antiangiogenic agents, antimetabolites,boron-containing agents, chemoprotective agents, hormones, antihormoneagents, corticosteroids, photoactive therapeutic agents,oligonucleotides, radionuclide agents, topoisomerase inhibitors,tyrosine kinase inhibitors, or radiosensitizers, as described in moreherein.

In a particular embodiment, the anti-CDCP1 antibodies disclosed hereinare used in combination with an anti-cancer agent or an antineoplasticagent. The terms “anti-cancer agent” and “antineoplastic agent” refer todrugs used to treat malignancies, such as cancerous growths. In oneembodiment, the anti-CDCP1 antibodies or ADCs of the invention areadministered in combination with one or more immune checkpointinhibitors (e.g., antibody or small molecule immune checkpointinhibitors) for the treatment of a cancer. In some embodiments, theimmune checkpoint inhibitor is an inhibitor of Programmed Death-Ligand 1(PD-L1, also known as B7-H1, CD274), Programmed Death 1 (PD-1), CTLA-4,PD-L2 (B7-DC, CD273), LAG3, TIM3, 2B4, A2aR, B7H1, B7H3, B7H4, BTLA,CD2, CD27, CD28, CD30, CD40, CD70, CD80, CD86, CD137, CD160, CD226,CD276, DR3, GAL9, GITR, HAVCR2, HVEM, IDO1, IDO2, ICOS (inducible T cellcostimulator), KIR, LAIR1, LIGHT, MARCO (macrophage receptor withcollageneous structure), PS (phosphatidylserine), OX-40, SLAM, TIGHT,VISTA, or any combinations thereof. In some embodiments, the immunecheckpoint inhibitor is an anti-CTLA-4, PD-L1, or PD-1 antibody therapysuch as, but not limited to Yervoy® (ipilimumab: Bristol-Myers Squibb),Opdivo® (nivolumab; Bristol-Myers Squibb), Keytruda® (pembrolizumabMerck), and Tecentriq® (atezolizumab; Roche).

In some embodiments, the anti-CDCP1 antibodies disclosed herein are usedin combination with an inhibitor of the PI3K-AKT mTOR pathway, e.g.,rapamycin, temsirolimus, everolimus, ridaforolimus, Torinl, Torin2,PP242, KU63794, WYE354, NVP-BEZ235, XL765, GDC-0491, GDC-0980,GSK2126458, AZD8055, OST-027, CH5132799, PF-05212384, or ZSTK474. Seealso WO 2012/054748; U.S. Pat. No. 8,394,818; McCubrcy et al., 2012,Oncotargct, 3:1068-1111.

In one embodiment, the anti-CDCP1 antibodies, bispecific antibodies, orADCs disclosed herein are administered in combination with a PARP (polyADP ribose polymerase) inhibitor. PARP inhibitors are well known tothose of ordinary skill in the art and include, but are not limited to,Niraparib, Olaparib, Rucaparib, Iniparib, Talazoparib, Veliparib, CEP9722, E7016, BGB-290, and 3-aminobenazamine.

Drug therapy may be used alone, or in combination with other treatmentssuch as surgery or radiation therapy. Several classes of drugs may beused in cancer treatment, depending on the nature of the organ involved.For example, breast cancers are commonly stimulated by estrogens, andmay be treated with drugs which inactive the sex hormones. Similarly,prostate cancer may be treated with drugs that inactivate androgens, themale sex hormone.

In particular embodiments, the anti-CDCP1 antibodies or ADCs can beadministered alone or with another anti-cancer agent which acts inconjunction with or synergistically with the antibody to treat thedisease associated with CDCP1 activity. Such anti-cancer agents include,for example, agents well known in the art (e.g., cytotoxins,chemotherapeutic agents, small molecules and radiation). Examples ofanti-cancer agents include, but are not limited to, Panorex(Glaxo-Welcome), Rituxan (IDEC/Genentech/Hoffman la Roche), Mylotarg(Wyeth), Campath (Millennium), Zevalin (IDEC and Schering AG), Bexxar(Corixa/GSK), Erbitux (Imclone/BMS), Avastin (Genentech) and Herceptin(Genentech/Hoffman la Roche). Other anti-cancer agents include, but arenot limited to, those disclosed in U.S. Pat. No. 7,598,028 andInternational Publication No. WO2008/100624, the contents of which arehereby incorporated by reference. One or more anti-cancer agents may beadministered either simultaneously or before or after administration ofan antibody or antigen binding portion thereof.

In particular embodiments of the invention, the anti-CDCP1 antibodies orADCs described herein can be used in a combination therapy with aninhibitor of NAMPT (see examples of inhibitors in US 2013/0303509;AbbVic, Inc., incorporated by reference herein) to treat a subject inneed thereof. NAMPT (also known as pre-B-cell-colony-enhancing factor(PBEF) and visfatin) is an enzyme that catalyzes the phosphoribosylationof nicotinamide and is the rate-limiting enzyme in one of two pathwaysthat salvage NAD. In one embodiment, anti-CDCP1 antibodies and ADCsdescribed herein are administered in combination with a NAMPT inhibitorfor the treatment of cancer in a subject.

In particular embodiments, the anti-CDCP1 antibodies or ADCs describedherein can be used in a combination therapy with SN-38, which is theactive metabolite of the topoisomerase inhibitor irinotecan.

The pharmaceutical compositions may include a “therapeutically effectiveamount” or a “prophylactically effective amount” of an antibody orantibody portion. A “therapeutically effective amount” refers to anamount effective, at dosages and for periods of time necessary, toachieve the desired therapeutic result. A therapeutically effectiveamount of the antibody or antibody portion may be determined by a personskilled in the art and may vary according to factors such as the diseasestate, age, sex, and weight of the individual, and the ability of theantibody or antibody portion to elicit a desired response in theindividual. A therapeutically effective amount is also one in which anytoxic or detrimental effects of the antibody, or antibody portion, areoutweighed by the therapeutically beneficial effects. A“prophylactically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredprophylactic result. Typically, since a prophylactic dose is used insubjects prior to or at an earlier stage of disease, theprophylactically effective amount will be less than the therapeuticallyeffective amount.

Dosage regimens may be adjusted to provide the optimum desired response(e.g., a therapeutic or prophylactic response). For example, a singlebolus may be administered, several divided doses may be administeredover time or the dose may be proportionally reduced or increased asindicated by the exigencies of the therapeutic situation. It isespecially advantageous to formulate parenteral compositions in dosageunit form for case of administration and uniformity of dosage. Dosageunit form as used herein refers to physically discrete units suited asunitary dosages for the mammalian subjects to be treated; each unitcontaining a predetermined quantity of active compound calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms aredictated by and directly dependent on (a) the unique characteristics ofthe active compound and the particular therapeutic or prophylacticeffect to be achieved, and (b) the limitations inherent in the art ofcompounding such an active compound for the treatment of sensitivity inindividuals.

An exemplary, non-limiting range for a therapeutically orprophylactically effective amount of an ADC, an antibody or antibodyportion is 0.1-20 mg/kg, more preferably 1-10 mg/kg. It is to be notedthat dosage values may vary with the type and severity of the conditionto be alleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat dosage ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed composition.

In another aspect, this application provides a method for detecting thepresence of CDCP1 in a sample in vitro (e.g., a biological sample, suchas serum, plasma, tissue, biopsy). The subject method can be used todiagnose a disorder, e.g., a cancer. The method includes: (i) contactingthe sample or a control sample with the anti-CDCP1 antibody or fragmentthereof as described herein; and (ii) detecting formation of a complexbetween the anti-CDCP1 antibody or fragment thereof, and the sample orthe control sample, wherein a statistically significant change in theformation of the complex in the sample relative to the control sample isindicative of the presence of CDCP1 in the sample.

Given their ability to bind to human CDCP1, the anti-human CDCP1antibodies, or portions thereof, (as well as ADCs thereof) can be usedto detect human CDCP1 (e.g., in a biological sample, such as serum orplasma), using a conventional immunoassay, such as an enzyme linkedimmunosorbent assays (ELISA), an radioimmunoassay (RIA) or tissueimmunohistochemistry. In one aspect, the disclosure provides a methodfor detecting human CDCP1 in a biological sample comprising contacting abiological sample with an antibody, or antibody portion, and detectingeither the antibody (or antibody portion) bound to human CDCP1 orunbound antibody (or antibody portion), to thereby detect human CDCP1 inthe biological sample. The antibody is directly or indirectly labeledwith a detectable substance to facilitate detection of the bound orunbound antibody. Suitable detectable substances include variousenzymes, prosthetic groups, fluorescent materials, luminescent materialsand radioactive materials. Examples of suitable enzymes includehorseradish peroxidase, alkaline phosphatase, β-galactosidase, oracetylcholinesterase; examples of suitable prosthetic group complexesinclude streptavidin/biotin and avidin/biotin; examples of suitablefluorescent materials include umbelliferone, fluorescein, fluoresceinisothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; an example of a luminescent material includesluminol; and examples of suitable radioactive material include ³H, ¹⁴C,³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹³¹I, ¹⁷⁷Lu, ¹⁶⁶Ho, or ¹⁵³Sm.

Alternative to labeling the antibody, human CDCP1 can be assayed inbiological fluids by a competition immunoassay utilizing rhCDCP1standards labeled with a detectable substance and an unlabeledanti-human CDCP1 antibody. In this assay, the biological sample, thelabeled rhCDCP1 standards and the anti-human CDCP1 antibody are combinedand the amount of labeled rhCDCP1 standard bound to the unlabeledantibody is determined. The amount of human CDCP1 in the biologicalsample is inversely proportional to the amount of labeled rhCDCP1standard bound to the anti-CDCP1 antibody. Similarly, human CDCP1 canalso be assayed in biological fluids by a competition immunoassayutilizing rhCDCP1 standards labeled with a detectable substance and anunlabeled anti-human CDCP1 antibody.

In yet another aspect, this application provides a method for detectingthe presence of CDCP1 in vivo (e.g., in vivo imaging in a subject). Thesubject method can be used to diagnose a disorder, e.g., aCDCP1-associated disorder. The method includes: (i) administering theanti-CDCP1 antibody or fragment thereof as described herein to a subjector a control subject under conditions that allow binding of the antibodyor fragment to CDCP1; and (ii) detecting formation of a complex betweenthe antibody or fragment and CDCP1, wherein a statistically significantchange in the formation of the complex in the subject relative to thecontrol subject is indicative of the presence of CDCP1.

Diagnostics and Imaging

The antibodies of the present disclosure can also be used in variousdiagnostic, and imaging methods. For example, the antibodies of thedisclosure can be used in any known assay method, such competitivebinding assays, direct and indirect sandwich assays, andimmunoprecipitation assays (see, e.g., Zola, Monoclonal Antibodies: AManual of Techniques, pp. 147-158 (CRC Press, Inc., 1987). For use insuch methods, e.g., for use in in vitro assays, the antibodies can bedetectably labeled, e.g., with a fluorophore such as Fluoresceinisothiocyanate (FITC) or phycoerythrin or with an enzyme substrate, suchas a substrate for horse radish peroxidase, for easy detection.

As discussed herein, the antibodies of the disclosure also can be usedfor in vivo diagnostic assays and in vivo imaging. In someimplementations, the antibody is labeled with a radionucleotide (such as³H, ¹¹¹In, ¹⁴C, ³²P, or ¹²³I) so that the cells or tissue of interestcan be localized and/or imaged using immunoscintigraphy. Methods ofconjugating labels to an antibody are known in the art. In otherimplementations of the disclosure, antibodies disclosed herein need notbe labeled, and the presence thereof can be detected using a labeledantibody, which binds to the antibody.

For example, the disclosure provides methods of detecting a CDCP1protein in a sample (e.g., a sample containing mammalian cells, e.g., abiopsy sample) that include contacting a sample with an antibodydisclosed herein and detecting binding of the antibody to any CDCP1protein in the sample. Some implementations further include recordingthe detection or non-detection of CDCP1 protein (e.g., the presence, thedetection, the non-detection, and/or level of a CDCP1 protein) in theclinical records of a subject from whom the sample was obtained.

In some implementations, the clinical record is stored on acomputer-readable medium, e.g., a disc, tape, or computer memory. Someimplementations further include administering any one of the antibodiesdescribed herein to a subject identified as having detectable CDCP1protein or an elevated level of a CDCP1 protein (e.g., as compared to areference level, e.g., a level of a CDCP1 protein produced by anon-cancerous cell) in his or her sample. Some implementations furtherinclude performing further testing for the presence of cancer (e.g., anyof the methods for further testing for the presence of cancer describedherein) on a subject identified as having detectable CDCP1 protein or anelevated level of a CDCP1 protein. Additional examples of referencevalues are described herein.

Also provided are methods of imaging one or more cancer cells (e.g., acancer cell that overexpresses CDCP1 or a cancer cell characterized byhaving CDCP1 on the cell surface, e.g., breast cancer (e.g.,triple-negative breast cancer), carcinoid cancer, cervical cancer,endometrial cancer, glioma, head and neck cancer, liver cancer, lungcancer, lymphoma, melanoma, ovarian cancer, pancreatic cancer, prostatecancer, renal cancer, colorectal cancer, gastric cancer, testicularcancer, thyroid cancer, bladder cancer, urethral cancer, or hematologicmalignancy cell) in a subject (e.g., a subject in need thereof, e.g., asubject identified as being at risk for developing a cancer, a subjectsuspected of having a cancer, or a subject already diagnosed oridentified as having a cancer), that include administering to a subjectan antibody that is conjugated to a detectable label (e.g., any of theexamples of detectable labels described herein) and imaging the presenceof the cancer cell by detecting the detectable label in the subject. Insome implementations, the detectable label is a fluorophore, ametalloporphyrin, a paramagnetic metal, a superparamagnetic metal, amagnetic particle (e.g., 10-20 nm in diameter), a nitroxide stable freeradical, or ferrioxamine methane sulfonate, or a metal, e.g., gold.

The methods of detection and diagnosis can be performed on anybiological sample (e.g., a sample containing mammalian cells), e.g., asample from an individual having a cancer or suspected of having acancer. The biological sample can be, e.g., a biopsy (e.g., needlebiopsy), tissue section, or a bodily fluid (e.g., lung gavage, urine,saliva, blood, tears, semen, or breast milk).

The antibody can also be used as staining reagent in pathology,following techniques that are well known in the art.

Such detection of specific binding by the antibody to the sample (e.g.,detection of an antibody: sample complex) can be made by any knownmethod including, without limitation, western blotting analysis,immunohistochemistry (IHC) analysis, immunofluorescence (IF) analysis,flow cytometry analysis, FACS analysis, ELISA, and immunoprecipitation.See, generally, Immunological Methods, Vols. I and II (Lefkovits andPernis, eds., Academic Press, N Y, 1979 and 1981, herein incorporated byreference.

Cellular extracts of the foregoing biological samples may be prepared,either crude or partially (or entirely) purified, in accordance withstandard techniques, and used in the methods of the disclosure.Alternatively, biological samples comprising whole cells, e.g.circulating tumor cells (CTCs), can be utilized in assay formats such asimmunohistochemistry (IHC), flow cytometry (FC), and immunofluorescence(IF).

V. Pharmaceutical Compositions

The disclosure also provides pharmaceutical compositions comprising anantibody, or antigen binding portion thereof, or ADC and apharmaceutically acceptable carrier. The pharmaceutical compositionscomprising antibodies or ADCs are for use in, but not limited to,diagnosing, detecting, or monitoring a disorder, in preventing,treating, managing, or ameliorating of a disorder or one or moresymptoms thereof, and/or in research. In a specific embodiment, acomposition comprises one or more antibodies. In another embodiment, thepharmaceutical composition comprises one or more antibodies or ADCs andone or more prophylactic or therapeutic agents other than antibodies orADCs for treating a disorder in which CDCP1 activity is detrimental.Preferably, the prophylactic or therapeutic agents known to be usefulfor or having been or currently being used in the prevention, treatment,management, or amelioration of a disorder or one or more symptomsthereof. In accordance with these embodiments, the composition mayfurther comprise of a carrier, diluent or excipient.

The antibodies and antibody-portions or ADCs can be incorporated intopharmaceutical compositions suitable for administration to a subject.Typically, the pharmaceutical composition comprises an antibody orantibody portion and a pharmaceutically acceptable carrier. As usedherein, “pharmaceutically acceptable carrier” includes any and allsolvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like that arephysiologically compatible. Examples of pharmaceutically acceptablecarriers include one or more of water, saline, phosphate bufferedsaline, dextrose, glycerol, ethanol and the like, as well ascombinations thereof. In many cases, it will be preferable to includeisotonic agents, for example, sugars, polyalcohols such as mannitol,sorbitol, or sodium chloride in the composition. Pharmaceuticallyacceptable carriers may further comprise minor amounts of auxiliarysubstances such as wetting or emulsifying agents, preservatives orbuffers, which enhance the shelf life or effectiveness of the antibodyor antibody portion or ADC.

Various delivery systems are known and can be used to administer one ormore antibodies or ADCs or the combination of one or more antibodies anda prophylactic agent or therapeutic agent useful for preventing,managing, treating, or ameliorating a disorder or one or more symptomsthereof, e.g., encapsulation in liposomes, microparticles,microcapsules, recombinant cells capable of expressing the antibody orantibody fragment, receptor-mediated endocytosis (see, e.g., Wu and Wu,J. Biol. Chem. 262:4429-4432 (1987)), construction of a nucleic acid aspart of a retroviral or other vector, etc. Methods of administering aprophylactic or therapeutic agent include, but are not limited to,parenteral administration (e.g., intradermal, intramuscular,intraperitoneal, intravenous and subcutaneous), epidural administration,intratumoral administration, and mucosal administration (e.g.,intranasal and oral routes). In addition, pulmonary administration canbe employed, e.g., by use of an inhaler or nebulizer, and formulationwith an aerosolizing agent. See, e.g., U.S. Pat. Nos. 6,019,968,5,985,320, 5,985,309, 5,934, 272, 5,874,064, 5,855,913, 5,290, 540, and4,880,078; and PCT Publication Nos. WO 92/19244, WO 97/32572, WO97/44013, WO 98/31346, and WO 99/66903, each of which is incorporatedherein by reference their entireties. In one embodiment, an antibody,combination therapy, or a composition is administered using AlkermesAIR® pulmonary drug delivery technology (Alkermes, Inc., Cambridge,Mass.). In a specific embodiment, prophylactic or therapeutic agents areadministered intramuscularly, intravenously, intratumorally, orally,intranasally, pulmonary, or subcutaneously. The prophylactic ortherapeutic agents may be administered by any convenient route, forexample by infusion or bolus injection, by absorption through epithelialor mucocutaneous linings (e.g., oral mucosa, rectal and intestinalmucosa, etc.) and may be administered together with other biologicallyactive agents. Administration can be systemic or local.

In a specific embodiment, it may be desirable to administer theprophylactic or therapeutic agents locally to the area in need oftreatment; this may be achieved by, for example, and not by way oflimitation, local infusion, by injection, or by means of an implant,said implant being of a porous or non-porous material, includingmembranes and matrices, such as sialastic membranes, polymers, fibrousmatrices (e.g., Tissuel®), or collagen matrices. In one embodiment, aneffective amount of one or more antibodies antagonists is administeredlocally to the affected area to a subject to prevent, treat, manage,and/or ameliorate a disorder or a symptom thereof. In anotherembodiment, an effective amount of one or more antibodies isadministered locally to the affected area in combination with aneffective amount of one or more therapies (e.g., one or moreprophylactic or therapeutic agents) other than an antibody of a subjectto prevent, treat, manage, and/or ameliorate a disorder or one or moresymptoms thereof.

A pharmaceutical composition is formulated to be compatible with itsintended route of administration. Examples of routes of administrationinclude, but are not limited to, parenteral, e.g., intravenous,intradermal, subcutaneous, oral, intranasal (e.g., inhalation),transdermal (e.g., topical), transmucosal, and rectal administration. Ina specific embodiment, the composition is formulated in accordance withroutine procedures as a pharmaceutical composition adapted forintravenous, subcutaneous, intramuscular, oral, intranasal, or topicaladministration to human beings. Typically, compositions for intravenousadministration are solutions in sterile isotonic aqueous buffer. Wherenecessary, the composition may also include a solubilizing agent and alocal anesthetic such as lignocaine to ease pain at the site of theinjection.

If the method of the disclosure comprises oral administration,compositions can be formulated orally in the form of tablets, capsules,cachets, gel caps, solutions, suspensions, and the like. Tablets orcapsules can be prepared by conventional means with pharmaceuticallyacceptable excipients such as binding agents (e.g., pregelatinised maizestarch, polyvinylpyrrolidonc, or hydroxypropyl methylcellulose); fillers(e.g., lactose, microcrystalline cellulose, or calcium hydrogenphosphate); lubricants (e.g., magnesium stearate, talc, or silica);disintcgrants (e.g., potato starch or sodium starch glycolate); orwetting agents (e.g., sodium lauryl sulphate). The tablets may be coatedby methods well-known in the art. Liquid preparations for oraladministration may take the form of, but not limited to, solutions,syrups or suspensions, or they may be presented as a dry product forconstitution with water or other suitable vehicle before use. Suchliquid preparations may be prepared by conventional means withpharmaceutically acceptable additives such as suspending agents (e.g.,sorbitol syrup, cellulose derivatives, or hydrogenated edible fats);emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles(e.g., almond oil, oily esters, ethyl alcohol, or fractionated vegetableoils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates orsorbic acid). The preparations may also contain buffer salts, flavoring,coloring, and sweetening agents as appropriate. Preparations for oraladministration may be suitably formulated for slow release, controlledrelease, or sustained release of a prophylactic or therapeutic agent(s).

The method may comprise administration of a composition formulated forparenteral administration by injection (e.g., by bolus injection orcontinuous infusion). Formulations for injection may be presented inunit dosage form (e.g., in ampoules or in multi-dose containers) with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle (e.g., sterile pyrogen-free water)before use.

Generally, the ingredients of compositions are supplied eitherseparately or mixed together in unit dosage form, for example, as a drylyophilized powder or water free concentrate in a hermetically sealedcontainer such as an ampoule or sachette indicating the quantity ofactive agent. Where the mode of administration is infusion, compositioncan be dispensed with an infusion bottle containing sterilepharmaceutical grade water or saline. Where the mode of administrationis by injection, an ampoule of sterile water for injection or saline canbe provided so that the ingredients may be mixed prior toadministration.

In particular, the disclosure also provides that one or more of theprophylactic or therapeutic agents, or pharmaceutical compositions ispackaged in a hermetically sealed container such as an ampoule orsachette indicating the quantity of the agent. In one embodiment, one ormore of the prophylactic or therapeutic agents, or pharmaceuticalcompositions is supplied as a dry sterilized lyophilized powder or waterfree concentrate in a hermetically scaled container and can bereconstituted (e.g., with water or saline) to the appropriateconcentration for administration to a subject. The antibodies andantibody-portions or ADCs can be administered by a variety of methodsknown in the art, although for many therapeutic applications, thepreferred route/mode of administration is subcutaneous injection,intravenous injection or infusion. As will be appreciated by the skilledartisan, the route and/or mode of administration will vary dependingupon the desired results. In certain embodiments, the active compoundmay be prepared with a carrier that will protect the compound againstrapid release, such as a controlled release formulation, includingimplants, transdermal patches, and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Many methods for the preparationof such formulations are patented or generally known to those skilled inthe art. See, e.g., Sustained and Controlled Release Drug DeliverySystems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

It will be readily apparent to those skilled in the art that othersuitable modifications and adaptations of the methods of the inventiondescribed herein are obvious and may be made using suitable equivalentswithout departing from the scope of the invention or the embodimentsdisclosed herein. Having now described the invention in detail, the samewill be more clearly understood by reference to the following examples,which are included for purposes of illustration only and are notintended to be limiting.

EXAMPLES

The following examples are provided to illustrate, but not to limit, theinvention.

Example 1. Preparation of Monoclonal Antibodies

Monoclonal antibodies were obtained by immunization of rabbits withrabbit splenocytes expressing full-length CDCP1 or immunization of miceengineered with the capacity to produce fully human immunoglobulins withrecombinant extracellular domain of CDCP1. Additionally, antibody 41A10was partially humanized to produce antibody ATF3. Sequences of the 41A10antibodies are disclosed in Table 2.

TABLE 2 Ab Isotype VH DNA VH VL DNA VL 41A10 Rabbit CAGGAGCAGCTGQEQLEESG GCATTCGAGTTG AFELTQTPA IgG GAGGAGTCCGG GDLVKPGA ACCCAGACTCCASVEADVGG GGGAGACCTGGT SLTLTCKSS GCCTCCGTGGAG TVTIKCQAS CAAGCCTGGGGCGIDFSSAYY GCAGATGTGGGA QNIYSNLA ATCCCTGACACT MCWVRQA GGCACAGTCACCWYQQKPGQ CACCTGCAAGTC PGKGLEWI ATCAAGTGCCAG PPKLLIYGA CTCTGGAATCGAACIYTGSTY GCCAGTCAGAAC STLASGVSS CTTCAGCAGTGC YANWAKG ATTTACAGCAATRFRGSGSGT CTATTACATGTG RFTISKTSS TTAGCCTGGTATC EFTLTISDLE CTGGGTCCGCCATTVTLQMT AACAGAAACCAG CADAATYY GGCTCCCGGGAA SLTAADTA GGCAGCCTCCCACQGGDDDS GGGGCTGGAGTG TYFCARDPI AGCTCCTGATCT YAFGGGTE GATCGCATGCATGYMFDLW ATGGTGCATCCA VVVK (SEQ TTATACTGGTAG GPGTLLTV CTCTGGCATCTGID NO: 405) CACTTACTACGC SS (SEQ  GGGTCTCATCGC GAACTGGGCGA ID NO:GGTTCAGAGGCA AAGGCCGATTCA 403) GTGGATCTGGGA CCATCTCCAAAA CAGAGTTCACTCCCTCGTCGACCA TCACCATCAGCG CGGTGACTCTGC ACCTGGAGTGTG AAATGACCAGTCCCGATGCTGCCA TGACAGCCGCGG CTTACTACTGTCA ACACGGCCACTT GGGCGGTGATGAATTTTTGTGCCA TGATAGTTATGCT GAGATCCTATTG TTCGGCGGAGGG GTTATATGTTTGACCGAGGTGGTG ACTTGTGGGGCC GTCAAAG CAGGCACCCTAC (SEQ ID NO: TCACCGTCTCCT404) CA (SEQ ID NO: 402)

Example 2. Generation of Antibodies Against the CDCP1 ExtracellularDomain

Experiments were performed to generate fully human antibodies againstCDCP1 extracellular domain. The following methods were used in theexamples.

Recombinant CDCP1 Cloning

Human CDCP1 cDNA was purchased from Origene (RC220633, Rockville, Md.)and named BBP374. The encoded protein was aligned with GenBankCDCP1_HUMAN sequence and found to differ at two positions: Q525R andD709G.

Ectodomains of human, cynomolgus monkey, rat, and mouse were cloned byeither PCR (human) or synthetic genes (monkey, rat, mouse). Thesynthetic genes were based on GenBank sequences (Table 3). All DNAsequences were cloned into appropriate CMV-based expression vectors withnon-native signal peptides and C-terminal histidine tags forpurification.

TABLE 3 Source of CDCP1 protein sequences Species GenBank ProteinReference Human CDCP1_HUMAN Cynomolgus monkey XP_005546930 RatNP_001100339 Mouse CDCP1_MOUSE

The Origene® BBP374 construct was used to express full-length humanCDCP1 protein (M1-E836) on the surface of HEK-293 cells. In addition, aseries of human CDCP1 constructs were generated to screen antibodybinding properties and epitopes (see Table 4).

TABLE 4 CDCP1 cell-surface expression vectors Plasmid name Sequencefeature Comment BBP374 M1-E836 Q525R D709G Native CDCP1 (plus Myc-DDK)pMSCV/FLCDCP1 M1-E836 Q525R D709G Native CDCP1 (plus Myc-DDKpMSCV/ClvCDCP1 K343-E836 Q525R D709G (N342) (plus Myc-DDK)

A series of CDCP1 constructs were generated to secrete portions of theectodomain from CHO cells. These plasmids were cloned (see Table 5) toassist with biophysical evaluations.

TABLE 5 CDCP1 ectodomain expression vectors Sequence feature Plasmid(plus 8xHis name Species (SEQ ID NO: 406)) Comment BBP476 Human F30-T667Full ectodomain BBP463 Human F30-T667 Q525R Full ectodomain with Q525RSNP BBP477 Human F30-T667 R368GS Impaired R368-K369 K369GS Q525Rcleavage BBP464 Human F30-R368 Distal domain BBP465 Human K343-T667Q525R Proximal domain BBP467 Human C221-E544 Q525R CUB1/CUB2 domainBBP468 Cynomolgus F30-T667 Full monkey ectodomain BBP469 Rat S30-T667Full rat ectodomain BBP470 Mouse S30-T663 Full mouse ectodomain

Cloning VH and VL Sequences from Hybridomas

For determination of CDR sequences, total RNA was isolated fromhybridoma cells using an RNeasy® kit (Qiagen, Hilden, Germany). Firstand second-strand cDNA synthesis was performed using a One Taq® One-StepRT-PCR kit (New England BioLabs, Ipswich, Mass.). PCR products wereseparated by agarose electrophoresis and fragments wcrc excised andpurified by a QlAquick® gel extraction kit (Qiagen, Hilden, Germany).Fragments were cloned directly into expression vectors with BspQI (NewEngland BioLabs, Ipswich, Mass.) by Golden Gate cloning techniques. Fourcolonies from each reaction were scaled up for miniprep-scale plasmidpurification by SequeMid® DNA Purification Kit (Aline Biosciences,Woburn, Mass.).

Identification of Functional, Recombinant VH and VL Sequences

For each hybridoma, four plasmids encoding the recombinant heavy chainwere paired with four plasmids encoding the recombinant light chain.These plasmid pairs were transfected into HEK-293 cells in 96-wellplates. Five days later conditioned medium from each pairing wasscreened by ELISA for binding to the target.

From each hybridoma there were 16 wells of transfected cells. Based onELISA data, one pairing (of the 16) was chosen from each hybridoma. Thecolonies containing plasmids that were used for the chosen pairing werecultured and plasmid DNA was purified at the maxiprep-scale (Qiagen).These plasmids were subjected to DNA sequence determination andanalysis.

Transient Expression System

The CDCP1 recombinant proteins and anti-CDCP1 antibodies were expressedin Chinese hamster ovary (CHO) cells using recommended transfection andmedia components of the ExpiCHO system (Invitrogcn, Carlsbad, Calif.).Cell culture supernatants were harvested 14 days post-transfection,centrifuged, and filtered (0.22 urn) prior to purification.

Purification of Recombinant His-Tagged Proteins

Conditioned medium from CHO cell cultures was clarified, filtered, andloaded onto an ÄKTA prime plus system with a 5 mL HisTrap™ FF column (GEHealthcare). Fractions were collected, analyzed by SDS-PAGE, pooled, anddialyzed against PBS.

Antibody Purification

Conditioned medium from CHO cell cultures was clarified, filtered, andpurified by loading onto an ÄKTA pure system with a 5 mL MabSelect SuRe®column (GE Healthcare). Antibodies were eluted with 100 mM glycine, pH3.5 and neutralized with 1M Tris-Cl, pH 8.5.

Recombinant Antibody Analyses

Concentration: Concentration of recombinant antibodies was determined ona Fortebio Octet using Protein A tips and a human IgG1 antibody for thestandard curve.

Purity testing by SDS-PAGE: Purity testing was performed by sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of reducedand non-reduced samples. Samples (10 ug) were mixed with loading buffer(+/−β-mercaptoethanol), heated, and electrophoresed on a 4-20% gel(Invitrogen). Bands were visualized by Coomassie InstantBlue® (Expedeon)staining.

Purity testing by Endotoxin: Endotoxin concentrations were measured bythe Limulus amoebocyte lysate (LAL) kinetic turbidometric method usingthe Endosafe-PTS® system (Charles River Laboratories).

Purity testing by HPLC-SEC: Samples were screened for aggregation orother forms of antibody on a 1260 Infinity System (Agilent) with aTSKgel UltraSW® Aggregate Guard column and HPLC column (TosohBioscience). Samples and standards were detected by absorbance at 280nm. Comparison against the standard curve provided the molar mass ofsample components.

Affinity: The affinity of antibodies to various recombinant CDCP1molecules was determined on an Octet Red (Pall, ForteBio) instrument.After loading reagents into a 96-well plate, the Octet Red with ProteinA-conjugated biosensors was programmed as follows: 30 seconds forbaseline #1; 120 seconds to immobilize the antibody; 30 seconds forbaseline #2; 300 seconds for association of antibody to recombinantCDCP1; and 300-600 seconds for dissociation of recombinant CDCP1 fromthe antibody.

Epitope binning: Binding competition among different antibodies wasdetermined using a real-time, interferometry assay on an Octet Red(Pall, ForteBio) instrument with Protein A-conjugated biosensors. Toassess whether two antibodies competed for binding to a recombinantCDCP1 protein, the assay was performed as follows. Protein A biosensorswere first submerged into wells containing 20 ug/mL of individualmonoclonal antibodies for 10 minutes. Following the capture step, thebiosensors were dipped briefly (30 sec) into buffer and then anyunoccupied sites on the biosensor were saturated by submerging them for5 minutes into wells containing 200 ug/mL of an irrelevant monoclonalantibody. The Octet biosensors were then dipped briefly (30 sec) inbuffer before immersion for 5 minutes into wells containing recombinantCDCP1. The biosensors were dipped briefly (30 sec) in buffer beforeimmersion for 5 minutes into wells containing a second recombinantantibody.

For the control case where the second antibody was the same as thefirst, there was no increase in signal, because there was no additionalbinding to the recombinant target. For the control case where buffer wasused instead of the first antibody, no recombinant target bound thenon-quenching antibody on the biosensor and no second antibody bound thebiosensor. For cases where a boost in signal was seen with the secondantibody, the two antibodies were determined not to compete. For caseswhere no boost in signal was seen with the second antibody, the twoantibodies were determined to compete for binding.

Immunofluorescence (IF) Based High Content Screening (HCS)

High content immunofluorescence was used to identify wells that containimmunoglobulin that preferentially bound CDCP1. Briefly, HCT116 cells(CDCP1+) and MCF7 cells (CDCP1−) seeded 24 hours before the assay wereincubated for 45 minutes at 37° C. with hybridoma supernatant diluted2-fold in DMEM+10% fetal bovine serum (FBS). After incubation, cellswere fixed in 4% formaldehyde, washed with PBS, permeabilized with 0.3%Triton®-X-100, and labeled with anti-human Alexa® 488 secondaryantibodies for 1 hour at room temperature. Unbound secondary antibodywas removed with PBS washes, and cells were stained with DNA dye(propidium iodide and Hoechst 33342).

Potential hits were initially identified via low-resolution, highthroughput screening using a TTP Labtech Acumen eX3® (TTP Labtech,Cambridge, Mass.), quantifying the fluorescence differential for eachsample on both positive and negative cell lines. Those hits weresubsequently verified and the subcellular localization of each samplewas characterized using a Thermo ArrayScan VTi® (Thermo FisherScientific, Waltham, Mass.) to obtain high-resolution images of bothcell lines.

Wells containing immunoglobulin that preferentially bound the CDCP1 wereanalyzed again on 293T cells or 293T cells over-expressing full length(FL) CDCP1 and cleaved-CDCP1 to confirm CDCP1 specificity and todetermine epitope localization.

Determination of Epitope Localization of CDCP1 Antibodies

Two proteolytic cleavage sites (R368 and N342) in the CDCP1 ECD wereidentified using mass spectrometry technology. 293T cells stablytransfected with DNA constructs expressing full length (FL) CDCP1 andcleaved (Clv) CDCP1 (AA343-836) expresses these proteins on the cellsurface. As shown in FIG. 1B, Ab1 binds to FL-CDCP1 only, therefore itsepitope is likely to be in the distal region (AA30-342). Ab2 binds toboth 293T/FL-CDCP1 and 293T/Clv-CDCP1, therefore its epitope is likelyto be in the proximal region (AA343-667). Antibodies that can bind both293T/FL-CDCP1 and 293T/Clv-CDCP1 (AA343-836) have epitopes localized inthe proximal region of CDCP1 ECD (AA343-667). In contrast, antibodiesthat can only bind 293T/FL-CDCP1 have epitopes localized to the distalregion of CDCP1 ECD (AA30-342) (FIGS. 1A-1C).

Results

Fully human antibodies against CDCP1 extracellular domain (ECD) weregenerated by standard hybridoma procedures. Briefly, mice were immunizedwith recombinant full length CDCP1 ECD or a cleaved form of ECD.Splenocytes were fused with the mouse myeloma cell line X63-Ag8.653.Clones producing antibodies against CDCP1 ECD were identified byimmunofluorescence (IF) based high content screening (HCS) on HCT116cells endogenously expressing CDCP1 and MCF7 cells not expressing CDCP1.CDCP1 specificity as well as ECD binding region was then determined byIF-HCS on 293T cells over-expressing full length (FL) CDCP1 andcleaved-CDCP1 (N342). The resulting CDCP1 specific clonal hybridomaswere cryopreserved in freezing medium and stored in liquid nitrogen.

Forty hybridoma hits were selected and successfully converted torecombinant human IgG1 antibody against CDCP1 including 21 Abs bindingto distal region (AA30-342) and 17 Abs binding to proximal region(AA343-667). Biological characterization of these antibodies includetarget internalization/degradation capabilities and matripase-mediatedR368 cleavage blockade. Biophysical characterization of these antibodiesinclude: cross-reactivity against human and Cynomolgus CDCP1, epitopecompetition binning, aggregation, degradation, stickiness and sequenceliability.

Eighteen recombinant monoclonal human antibodies were selected forepitope binning assay, kinetic analysis against human and CynomolgusCDCP1 (FIG. 15 ). The properties of these antibodies are summarized inFIG. 16 and FIG. 17 , and the sequences of these antibodies are shown inFIG. 18 , FIG. 19 , and FIG. 20 .

FIG. 16 and FIG. 17 . show the affinity of the antibodies to CDCP1.Affinity was measured by dissociation constant (K_(a)). K_(d) is theratio of the rate constant of the off rate (k_(off)) divided by the rateconstant of the on rate (k_(on)). The lower the value of K_(d), thehigher affinity of the antibody for the target molecule. FIG. 16 showsK_(d) for cynomolgus monkey CDCP1 and affinity to murine CDCP1. FIG. 17shows K_(d) for human CDCP1, K_(d) for cynomolgus monkey CDCP1, andaffinity to murine CDCP1. FIG. 18 shows the amino acid sequence of theheavy chain variable region (VH) and the light chain variable region(VL) of the 40 anti-CDCP1 antibodies. FIG. 19 shows the nucleotidesequence of the heavy chain variable region and the light chain variableregion of these antibodies. FIG. 20 shows the amino acid sequence of thecomplementary determining regions in the heavy chain variable region andcomplementary determining regions in the light chain variable region ofthe 40 anti-CDCP1 antibodies.

Example 3. CDCP1 is Highly Expressed and Phosphorylated in MultipleSolid Tumors

Experiments were performed to determine CDCP1 expression andphosphorylation in multiple solid tumors. The following methods wereused in this example.

Phospho-Tyrosine Peptide Profiling by PhosphoScan™

Tissue specimens were taken intra-operatively and snap frozen in liquidnitrogen. An average of 15 milligrams of peptides were prepared from0.2-0.5 grams of resected frozen ovarian tissues by homogenization,trypsin digestion and Sep-pak® C18 column purification. The methods aredescribed, e.g., in, Rush, John, et al. “Immunoaffinity profiling oftyrosine phosphorylation in cancer cells,” Nature Biotechnology, 23.1(2005): 94-101; and Gu, Ting-Lei, et al. “Survey of tyrosine kinasesignaling reveals ROS kinase fusions in human cholangiocarcinoma,” PloSone 6.1 (2011): e15640.

Peptides containing phospho-tyrosine (pY) were isolated byimmuno-precipitation with a monoclonal antibody against phospho-tyrosine(pY100), concentrated on reverse-phase micro tips, and analyzed byliquid chromatography, tandem mass spectrometry (LC-MS/MS). Briefly,samples were collected with an LTQ-Orbitrap™ mass spectrometer, using atop-ten method, a dynamic exclusion repeat count of 1, and a repeatduration of 30 seconds. MS and MS/MS spectra were collected in theOrbitrap and LTQ component of the mass spectrometer, respectively.SORCERER-SEQUEST™ (v4.0.3 (c) 2008, Sage-N Research, Inc., Milpitas,Calif.) searches were done against the NCBI human RefPept databasedownloaded on Jan. 6, 2009 (containing 37,742 proteins) or Mar. 1, 2010(containing 36,500 proteins), allowing for serine, threonine andtyrosine phosphorylation (STY+80) and methionine oxidation (M+16) asdifferential modifications.

Clustering and Ranking Analysis

To assess potentially aberrant tyrosine phosphorylation of proteins intumor tissues, spectral counts per protein were summed and normalized tothe amount of peptide subjected to pY immuno-precipitation (15 mg).Elevated spectral count in each tumor sample was calculated bysubtracting an average spectral count in normal tissues. Elevated pYspectral count proteins, representing elevated tyrosine phosphorylation,observed in tumor samples were used as the basis for hierarchicalclustering using the Pearson correlation distance metric and averagelinkage (MultiExperiment Viewer version 4.4). Proteins with elevatedphosphorylation in tumors were ranked based on the average value ofelevated spectral count in corresponding tumors. GenePattern 3.0software package (Broad Institute of MIT and Harvard, Cambridge, Mass.)was used for Comparative Marker Selection analysis.

Results

Using an unbiased and global phospho-proteomic approach, thePhosphoScan® program, CDCP1 was found to be differentially expressed inbreast, colon, lung, pancreatic, ovarian, and kidney cancer as comparedto normal tissue. Additionally, differential CDCP1 tyrosinephosphorylation was detected in breast, colon, lung, and ovarian cancer.CDCP1 is among the most differentially phosphorylated proteinsidentified in breast cancer, non-small cell lung cancer, liver cancerand colon cancer. Particularly, CDCP1 is highly phosphorylated inmalignant breast tumor tissue when compared with benign breast tumor.These results show that CDCP1 is highly expressed in multiple solidtumors and is a suitable target for cancer treatment using the ADCsdescribed herein.

Example 4. CDCP1 is Overexpressed in Triple Negative Breast Cancer(TNBC) and Colorectal Cancer (CRC)

Experiments were performed to determine CDCP1 expression in TNBC andCRC. The following methods were used in this example.

4 μm FFPE tissue sections or tissue microarrays (TMA) slides werede-paraffinized and rehydrated through xylene and graded ethanol,respectively. Antigen retrieval was performed in a Decloaking Chamber(Biocare Medical, Concord, Calif.) using 1.0 mM EDTA, pH 8.0. Slideswere then quenched in 3% H₂O₂ for 10 minutes, washed in deionized H₂Oand blocked with Tris buffered saline/0.5% Tween-20 (TBST)/5% goat serumin a humidified chamber for 60 minutes. Sections were then exposed toanti-CDCP1 antibody overnight at 4° C. Detection was performed withSignalStain® Boost IHC Detection Reagent (Cell Signaling Technology,Danvers, Mass.) for 30 minutes. All slides were exposed to NovaRed®(Vector Laboratories, Inc., Burlingame, Calif.) for 1 minute before theywere rinsed, dehydrated, cleared and cover-slipped. Animmunohistochemistry (IHC) score was given to each specimen using aqualitative scoring method. The scoring method categorizes differentlevels of IHC staining into different groups, including negative (−),weak (1+), moderate (2+), and strong (3+ or more). The method ofdetermining the IHC scores is described in, e.g., Fedchenko, Nickolay,and Janin Reifenrath. “Different approaches for interpretation andreporting of immunohistochemistry analysis results in the bone tissue—areview.” Diagnos. Pathol., 9 (1): 221 (2014), which is incorporatedherein by reference in its entirety.

Immunohistochemistry (IHC) analysis further revealed 86% of 26 primaryTNBC tumor tissue over-express CDCP1 (IHC score of 2+ or more). Intumor, CDCP1 is localized predominantly on the cell surface. 7/7metastatic lymph node specimens from TNBC patients overexpressed CDCP1(IHC score of 2+ or more). In addition, 2 relapsed TNBC tumorsoverexpressed CDCP1.

In colon canccr, 12 primary tumor specimen and 4 metastatic specimen ofcolorectal cancer harvested from liver, lung or pancreas exhibited highlevel of membrane expression of CDCP1 (Table 6).

TABLE 6 Overexpression of CDCP1 in Primary, Metastatic and/or RelapsedTumors of TNBC and CRC Total IHC IHC IHC IHC ≥ # of tissue IndicationTissue Type No. 0 1+ 2+ 3+ IHC ≥ 2+ TNBC Primary 26 0 4 12 12 24 (86%) tumor TNBC Normal 5 5 0 (0%)  adjacent tissue TNBC Lymph node 7 3 4  7(100%) metastatic tumor TNBC Relapsed 2 1 1  2 (100%) tumor CRC Primary12 12 12 (100%) tumor CRC Metastatic 4 4  4 (100%) tumor (liver, lung,pancreas)

These results confirm that CDCP1 is overexpressed in primary,metastatic, and relapsed tumors, which are thus suitable targets forcancer therapy using the ADCs described herein.

Example 5. CDCP1 is Expressed in Other Subtypes of Breast Cancer

Immunohistochemistry (IHC) analysis revealed that CDCP1 is overexpressedin 5/5 ductal carcinoma in situ (DCIS), 5/5 luminal A breast cancer, 4/4Her2+ breast cancer tumor specimens (Table 7). In addition, 2 tissuemicroarray containing 138 and 149 scorable tumor tissue cores frominvasive ductal carcinoma (IDC) patients with known Her2 expressionstatus were examined for CDCP1 expression by immunohistochemistryanalysis. High level of CDCP1 expression was detected in 46-57% of Her 2over-expressing tumor specimen versus 18% or 54% of Her 2 low tumorspecimens (Table 7).

TABLE 7 Overexpression of CDCP1 in several subtypes of breast cancerspecimen Total IHC IHC IHC IHC ≥ # of tissue Type of BrCa specimen No. 01+ 2+ 3+ IHC ≥ 2+ DCIS 5 0 0 3 2 5 (100%) Luminal A 5 0 0 2 3 5 (100%)Her2+ breast cancer 4 0 0 2 2 4 (100%) IDC tissue microarrays 66 39 16 92 11 (18%)  (TMA-1) (Her2 expression IHC 0-1+) IDC tissue microarrays 7219 20 23 10 33 (46%)  (TMA-1) (Her2 expression IHC 2-3+) IDC tissuemicroarrays 81 11 26 18 26 44 (54%)  (TMA-2) (Her2 expression IHC 0-1+)IDC tissue microarrays 68 11 13 15 24 39 (57%)  (TMA-2) (Her2 expressionIHC 2-3+)

Example 6. CDCP1 is Expressed in Other Types of Solid Tumors

Immunohistochemistry (IHC) analysis revealed that CDCP1 is overexpressedin other types of solid tumors. In ovarian serous carcinoma, CDCP1overexpression is detected in 95% of the 18 primary tumors and 72% ofthe 11 relapsed tumors. In ovarian clear cell carcinoma, CDCP1 isoverexpressed in 5/5 samples. Among 36 Non-Small Cell Lung Cancer(NSCLC), 8 pancreatic cancer and 14 prostate cancer specimens, 86%, 100%and 79% of them overexpressed CDCP1, respectively. 20% of the 20 SmallCell Lung Cancer (SCLC) specimen overexpressed CDCP1 (Table 8).

TABLE 8 Overexpression of CDCP1 in other types of solid tumors Tissue #of tissue Indication Type Total 0 1+ 2+ ≥3+ IHC ≥ 2+ Ovarian Primary 180 1 5 12 17 (95%)  serous tumor carcinoma Recurrent Relapsed 11 1 2 4 48 (72%)  serous carcinoma Ovarian Primary 5 0 0 0 5 5 (100%) clear celltumor carcinoma NSCLC Primary 36 3 2 10 21 31 (86%)  tumor PancreaticPrimary 8 0 0 3 5 8 (100%) Cancer tumor Prostate Primary 14 0 3 3 8 11(79%)  Cancer tumor SCLC Primary 20 8 8 2 2 4 (20%)  tumor

Example 7. CDCP1 is Expressed in TNBC, Her2+ Breast Cancer and CRC CellLines

A panel of TNBC, Her2+ breast cancer and CRC cell lines were examinedfor their surface CDCP1 expression using a mouse monoclonal anti-CDCP1antibody directly conjugated to fluorochrome phycoerythrin (PE)(Biolegend, San Diego, Calif., Cat. No. 324006). Antigen density wasdetermined using BD Quantibrite™ Beads PE Fluorescence Quantitation Kit(BD Cat. No. 340495).

Flow cytometry analysis was performed for five TNBC cell lines MDA-231,BT549, DU4475, MDA-468 and HCC1187 using a mouse monoclonal anti-CDCP1antibody directly conjugated to fluorochrome PE. A breast cell line MCF7was used as a negative control. The estimated cell surface CDCP1 densitywas 89439 for MDA231, 202780 for BT549, 67543 for DU4475, 34340 forMDA468, 20255 for HCC1187, and 0 for MCF7 (negative control). Thus, cellsurface CDCP1 density was estimated at 20255-202780 for tested TNBC celllines.

Flow cytometry analysis was also performed for two Her2+ breast cancercell lines BT474 and SKBR3 using a mouse monoclonal anti-CDCP1 antibodydirectly conjugated to fluorochrome PE. A breast cell line MCF7 was usedas a negative control. The estimated cell surface CDCP1 density was67543 for BT474, 1101 for SKBR3, and 0 for MCF7 (negative control).Thus, cell surface CDCP1 density was estimated at 1101-67543 for testedHer2+ breast cancer cell lines.

Furthermore, flow cytometry analysis was performed for three colorectalcancer cell lines SW48, HCT116 and HCT8, using a mouse monoclonalanti-CDCP1 antibody directly conjugated to fluorochromc PE. Theestimated cell surface CDCP1 density was 69161 for SW48, 99921 forHCT116, and 25118 for HCT8. Thus, cell surface CDCP1 density wasestimated at 25100-99900 for tested CRC cells.

These results show that CDCP1 is present on the cell surface of cellsfrom various cancer cell lines, and suggest that the ADCs describedherein that target CDCP1 can be used to treat different types ofcancers.

Example 8. In Vitro Target Internalization/Degradation Assay

After confirming the target specificity of an antibody, the ability ofan antibody to induce CDCP1 internalization and degradation is assessedby incubating test antibodies at ˜10 ug/ml with live HCT116 cells platedin 384 well plates for 30 minutes, 2 hours, and overnight. Cells werethen fixed with 4% formaldehyde in PBS at room temperature for 15minutes, the plates were then washed three times with PBS, and blockedwith 5% normal goat scrum in PBS-T, mouse monoclonal antibody CUB1 wasthen added in Ab dilution buffer at 0.5 ug/ml to the wells. The plateswere then incubated at 4° C. overnight. On the next day, plates weredeveloped with anti-human IgG A488 and anti-mouse IgG A555 secondaryantibodies and imaged using an array scanner.

The live cell immunofluorescence assay was performed with two humanmonoclonal antibodies, 27H10 and 38E11. 27H10 induces CDCP1internalization and degradation, whereas 38E11 binds to the cell surfaceCDCP1 and docs not induced target internalization and degradation. 19out of 40 recombinant antibodies (FIG. 16 ) which likely bind toproximal region (AA343-667) bind to surface CDCP1 without internalizingthe target, as indicated by membranous staining of both human antibodyand CUB1 antibody; while 21 out of 40 recombinant antibodies (FIG. 16 )which likely bind to distal region (AA30-342) can cause strongintracellular punctate staining as well as target degradation indicatedby diminished antibody signal at 2 hours and overnight time points.

In summary, this example shows that certain CDCP1 antibodies can induceCDCP1 internalization and degradation.

Example 9. Identification of Specific Epitope Bins in Both Distal andProximal Regions of CDCP1

Experiments were performed to identify specific epitope bins in bothdistal and proximal regions of CDCP1. The following methods were used inthe examples.

R368 Cleavage Blocking Assay

CDCP1 can be proteolytically cleaved by matriptase at the C-terminalside of R368. Antibodies that can block CDCP1 cleavage at 8368 caninhibit cancer cell metastasis in mouse models. To determine if any ofthe human antibodies generated can block this cleavage, recombinantCDCP1 ECD (AA30-667) with a C-terminal His tag were first incubated withthe antibodies described in the examples at a protein/antibody ratio of1:3 on ice for 1 hour, matriptase was then added to each reaction at S:Eratio of 50:1 and incubated at 37° C. for 1.5 hour. The reaction wasthen boiled in SDS loading buffer and analyzed by Western Blot usinganti-His tag antibody, which detects both FL and Clv ECD.

As shown in FIGS. 2A-2C, some human antibodies can block matriptasecleavage of CDCP1, indicating that these antibodies are binding epitopesnear the R368 cleavage site and interfere with matriptase binding. 8 outof 20 recombinant human antibodies were found to be 8368 cleavageblockers. Therefore, these antibodies are in a special epitope bin thatmost likely resides in the proximal region (AA343-667).

Identification of N342 Clv-CDCP1 Preferred Antibodies

Two of the antibodies bind better to Clv-CDCP1 (N342) than to FL-CDCP1,indicating that these antibodies might bind to special epitopes enrichedor specific to Clv-CDCP1 (N342). Both an immunofluorescence assay using293T cells overexpressing FL and Clv CDCP1 (N342) as well as an Octetkinetics assay using recombinant FL-ECD and Clv-ECD (N342) demonstratedClv-CDCP1 preference of these two antibodies (FIG. 3 ). Sequenceanalysis of these two antibodies, 18C6 and 11F9, revealed that theybelong to the same VDJ family, which indicates that they most likelybind to the same epitope.

Results

Combining epitope binning results from Octet and biological analysis ofrecombinant monoclonal human antibodies, six distinct epitope bins wereidentified (FIG. 4 ) including two in the distal region (AA30-342): DB1and DB2; three in the proximal region (AA343-667): PB1, PB2 and PB3; andone N342 cleaved form preferred bin: CB1.

Example 10. In Vitro Target Modulation in Cultured Cancer Cell Lines

In addition to target internalization and degradation, other aspects oftarget modulation by the disclosed antibodies were assessed in severalcell based assays. TNBC cell line BT549 and CRC cell line SW48 wereseeded in 6 well plates at ˜70% confluency and were incubated with 5ug/ml recombinant human monoclonal antibodies against CDCP1 diluted incomplete culture medium for 0, 0.5, 1, 4, 6 hours and overnight. Cellswere then harvested in SDS-loading buffer and subjected to western blotanalysis using the following antibodies: anti-CDCP1 (Cat. No. 4115, CellSignaling Technology, Danvers, Mass.), anti-phosphoCDCP1 (pY734) (Cat.No. 9050, Cell Signaling Technology, Danvers, Mass.), anti-PKCδ (Cat.No. 9616, Cell Signaling Technology, Danvers, Mass.), anti-PKCS (pY311)(Cat. No. 2055, Cell Signaling Technology, Danvers, Mass.), anti-Erk(Cat. No. 4695, Cell Signaling Technology, Danvers, Mass.) and anti-Erk(pT202/Y204) (Cat. No. 4370, Cell Signaling Technology, Danvers, Mass.).

Table 9 summarizes the results generated in BT549 and SW48 cells. Asshown in Table 9, in both BT549 and SW48 cells, antibodies that caninternalize CDCP1 in immunofluorescence assay can induce CDCP1phosphorylation and PKC phosphorylation, indicating that theseantibodies act as CDCP1 agonists. Consistent with results from theimmunofluorescence analysis, these antibodies also caused subsequentCDCP1 degradation at later time points. In contrast, the threeantibodies that do not induce CDCP1 internalization in theimmunofluorescence assay did not increase CDCP1 phosphorylation or PKCδphosphorylation.

TABLE 9 In vitro target modulation and signaling effects of human CDCP1antibodies In vitro CDCP1 Epitope internal- p-CDCP1 degrada- p-PKCD Ablocalization ization (Y734) tion (Y311) 03B11 Distal Yes Yes Yes Yes(AA30-342) 27H10 Distal Yes Yes Yes Yes (AA30-342) 47G7 Distal Yes YesYes Yes (AA30-342) 41A9 Proximal No No No No (AA343-667) 38E11 ProximalNo No No No (AA343-667) 18C06 Proximal No No No No (AA343-667)

Example 11. Conjugation of Human Antibodies to vcMMAE

Human monoclonal antibodies against CDCP1 were conjugated to vcMMAE. Themethods were described, e.g., in Doronina, Svetlana O., et al.“Development of potent monoclonal antibody auristatin conjugates forcancer therapy.” Nature biotechnology 21.7 (2003): 778-784; andFrancisco, Joseph A., et al. “cAC10-vcMMAE, an anti-CD30-monomethylauristatin E conjugate with potent and selective antitumor activity.”Blood 102.4 (2003): 1458-1465.

Each antibody was mixed with dithiothreitol (DTT) at room temperaturefor 30 minutes, and the buffer was exchanged by Amicon spin columns (30kDa) into PBS pH 7.0 with 2 mM EDTA. The antibody concentration wasquantified by A280. They were then diluted to 1.5 mg/ml in PBS pH 7.0and 2 mM EDTA. A 4-fold molar excess of maleimidocaproyl-Val-Cit-MMAE(vc-MMAE) was then added to the reduced antibody at room temperature for1 hour. The reaction mixture was then dialyzed against 1×PBS pH 8.0 andfilter sterilized. The drug:mAh ratio (DAR) was then calculated based onspec readings at A280 and A248 on an absorption spectroscopy. The methodof calculating DAR is described in detail, e.g., in Hamblett, Kevin J.,et al. “Effects of drug loading on the antitumor activity of amonoclonal antibody drug conjugate,” Clinical Cancer Research 10.20(2004): 7063-7070.

Example 12. In Vitro Cancer Cell Killing Assay

Human antibodies conjugated to vc-MMAE were tested for their effect ongrowing TNBC and CRC cancer cells.

TNBC cell lines, BT549, MDA-MB-231, which express CDCP1, breast cancercell line MCF7, which does not express CDCP1, and CRC cell lines, SW48and HCT116, which express CDCP1, were seeded onto 96 well plate at2000-4000 cells/well. 24 hours later, Ab-vcMMAE bearing a drug antibodyratio (DAR) of 3.7-4.3 was added to the wells in complete culture mediumso that the final Ab-vcMMAE concentration in the well is 0.001, 0.01,0.1 and 1.0 ug/ml. Each treatment was replicated in 2-3 wells. 72 hourslater, cell viability was measured by CellTiter Glo® Luminescent Cellviability assay (Promega, Madison, Wis.) according to manufacturer'sinstructions. Cell viability was graphed by Prism® software using ratioof cell viability of test conditions to that of control wells that wereincubated with growth medium.

As shown in FIGS. 5A-5E, while none of the human Ab-vcMMAE could killMCF7, which does not express CDCP1, they showed varying degrees ofkilling effects on BT549, MDA231, SW48, and HCT116 cells. Noticeably,antibodies that can induce internalization in the immunofluorescenceassay (3B11-MMAE, 27H10-MMAE, and 47G7-MMAE) exhibited a better killingeffect (50-60% in BT549, 35-60% in SW48) than the ones (41A9-MMAE,38E11-MMAE, 18C6-MMAE) that do not (35% in BT549, 35-40% in SW48).

Example 13. In Vitro ADCC Assay

The capability of a few human antibodies of inducing antibody dependentcytotoxicity (ADCC) was tested using an ADCC Reporter Bioassay (Promega,Madison, Wis.). BT549 or SW48 cells were plated at 1×10⁴ cells per well24 hours in advance of treatment in sterile 96-well black cell culturemicroplate (GrenierBio, Kremsmünster, Austria). Culture medium wasaspirated in advance of treatment and 25u1 of RPMI 1640 supplementedwith 4% ultra-low IgG FBS was added to cells. Human monoclonalantibodies were diluted in assay medium and added to cells in 25 ul/wellin duplicate to achieve final antibody concentrations of 0, 0.0001,0.001, 0.01, 0.1, and 1.0 ug/ml. Engineered Jurkat effector cells werethawed at 37° C. for 2 minutes into assay medium and added to the cellsat an effector/target ratio of 7.5:1. The mixed cell culture wasincubated at 37° C., 5% CO₂ for 6 hours. Cell viability was analyzedusing Bio-Glo luciferase assay reagent as directed in technical manual.Relative Light Units (RLU) readout was graphed using Prism® software.

As shown in FIGS. 6A-6C, among tested antibodies, 41A9 was identified asthe antibody with highest ADCC activity in both TNBC cell lines, BT549and MDA231, as well as CRC cells SW48. 38E11 also exhibited ADCCactivity in these cell lines.

Example 14. In Vitro CDC Assay

The capability of six human antibodies, which bind to different epitopesin distal and proximal regions, of complement dependent cytotoxicity(CDC) is tested. BT549 or SW48 cells were plated at 1×10⁴ cells per wellin sterile 96-well black cell culture microplate (GrenierBio,Kremsmünster, Austria) in 50 ul RPMI 1640 supplemented with 5% heatinactivated ultra low IgG FBS (Life Technologies, Carlsbad, Calif.)assay medium. Human monoclonal antibodies were diluted in assay mediumand added to cells in 25 ul/well in duplicate and incubated at 37° C. 5%CO₂ for 15 minutes. Active and heat-inactivated baby rabbit complementwas added to experimental wells at a final concentration of 5%. Thefinal antibody concentrations of diluted antibodies are 0, 0.001, 0.01,0.1, 1.0, and 10.0 ug/ml. All wells normalized with assay medium to afinal volume of 100 ul and incubated at 37° C. 5% CO₂ for 6 hours. Cellviability was analyzed using Cell Titer Glo 2.0 (Promega, Madison, Wis.)assay reagent as directed in technical manual. Results were graphed aspercent of control wells using Prism® software. Ultra low IgG FBS forassay medium and negative control baby rabbit complement were heatinactivated at 56° C. for 30 minutes.

As shown in FIGS. 7A-7G, antibodies that can induce CDCP1internalization in cell culture, 3B11, 27H10, 47G7, induced a strongerCDC effect than those which do not. The dose-dependent CDC effect iscomplement dependent as indicated by inactivation of such activity whenheat inactivated baby rabbit complement (BRC) was used in the assay.

Example 15. In Vivo Antibody Tumor Penetration and Target Degradation

Mice with HCT116 xenograft tumors of 400-500 mm³ in size received 5mg/kg control antibodies (rabbit isotype control IgG or an anti-DDR1rabbit monoclonal antibody) or 41A10, a rabbit monoclonal antibodyagainst distal region of CDCP1, by tail intravenous injection. 4 and 24hours after i.v. injection, tumors were harvested and processed intoFFPE blocks. FFPE sections were then analyzed by an IHC antibody bindingto C-terminus of CDCP1. While tumors treated with isotype controlantibodies show strong and even CDCP1 staining, tumor treated with 41A10exhibited massive CDCP1 clearance. The target clearance can be seenaround blood vessel as early as 4 hours after i.v. injection, indicatingefficient tumor penetration and target degradation by 41A10 in this CRCtumor model. Thus, 41A10 (Rb) induced CDCP1 degradation in HCT116xenograft tumors.

Example 16. Conjugated CDCP1 Antibodies Cause Tumor Regression: TripleNegative Breast Cancer

Since the rabbit antibody 41A10 exhibited efficient tumor penetrationand target clearance in vivo, the ability of conjugated 41A10,41A10-vcMMAE was tested in a MDA231 xenografts, a TNBC tumor model.

MDA231 xenograft tumors were generated by injecting 2.5×10e6 MDA231cells in the presence of matrigel subcutaneously into 6-8 weeks oldfemale Ncr-nude mice (Taconic, Hudson, N.Y.). When most tumors reached200 mm³ in size (˜3 weeks after cell inoculation), mice are randomizedinto 4 treatment groups of 8-10 mice using an R studio script designedto group the animals so that the average tumor size and standarddeviation of the tumor size between groups are comparable.

The animals were then injected intravenously with 41A10 antibody, theantibodies conjugated to MMAE, a rabbit IgG isotype control antibody(No. 3900, Cell Signaling Technology, Danvers, Mass.) conjugated toMMAE, and PBS vehicle. ADCswere administered at 5 mg/kg for 4 doses overa period of 10 days, naked antibodies were administered at 10 mg/kg 2times a week until the end of the study (a total of 15 doses). Tumorvolume was estimated by 0.52×Length×width×width. As shown in FIGS. 8Aand 8B, while naked antibody 41A10 exhibited no effect on tumor growthin MDA-231 xenografts, 41A10-MMAE exhibited >150% tumor growthinhibition. Although 41A10 cross reacts with mouse CDCP1, 41A 10-MMAEtreated mice are healthy and active, as shown by the stable body weightand normal activities.

Example 17. Conjugated CDCP1 Antibodies Cause Tumor Regression: ColonCarcinoma

The human CDCP1 antibodies conjugated MMAEs were tested to demonstratetheir effect on tumor growth in vivo in a model of colon carcinoma. CRCtumor model SW48 xenografts were generated by injecting 2.5×10⁶ SW48cells in the presence of matrigel subcutaneously into 6-8 weeks oldfemale Ncr-nude mice (Taconic). When most tumors reach 200 mm³ in size(˜3 weeks after cell inoculation), mice arc randomized into 8 treatmentgroups of 8-10 mice using an R studio script designed to group theanimals so that the average tumor size and standard deviation of thetumor size between groups are comparable. The animals were then injectedintravenously with PBS, human IgG1 (isotype control)-MMAE and CDCP1Ab-MMAEs that showed in vitro cell killing effect at 5 mg/kg. The i.v.treatment was repeated 3 more times over a period of 10 days. Followingtreatment, tumor volumes and mouse weights were monitored until tumorsexceed 800 mm³. Mice that showed a 15% or more weight loss or becamelethargic are taken out of the study.

As shown in FIG. 9 , all six human antibody MMAE conjugates exhibitedsignificant tumor growth inhibition (180-250%) in SW48 tumors.

Example 18. Antibody Drug Coniueates Cause Tumor Regression in TNBCTumor Models

The human CDCP1 antibodies 18C6, 27H10, 38E11 and 41A9 conjugated withvcMMAE were also tested to demonstrate their effect on tumor growth inTNBC tumor model. TNBC tumor model MDA231 xenografts were generated byinjecting 2.5×10⁶ corresponding cultured cells in the presence ofmatrigel subcutaneously into 6-8 weeks old female Ncr-nude mice(Taconic). When most tumors reach 250-300 mm³ in size (˜3 weeks aftercell inoculation), mice are randomized into treatment groups of 8-10mice using an R studio script designed to group the animals so that theaverage tumor size and standard deviation of the tumor size betweengroups are comparable.

The animals were then injected intravenously with vehicle, human IgG1(isotype control)-MMAE and 4 human CDCP1 ADCs (18C6-MMAE, 27H10-MMAE,38E11-MMAE and 41A9-MMAE) at 5 mg/kg. The i.v. treatment was repeated 3times over a period of 10 days. Following treatment, tumor volumes andmouse weights were monitored until tumors exceed 1500 mm³. Mice thatshowed a 15% or more weight loss or become lethargic were taken out ofthe study.

The four human antibody MMAE conjugates caused complete tumor regressionin the TNBC model MDA231 xenografts at 5 mg/kg (FIG. 10 ).

Example 19. Antibody Drug Conjugates Cause Tumor Regression in CRC TumorModels

The human CDCP1 antibodies 18C6, 27H10, 38E11 and 41A9 conjugated withvcMMAE were also tested to demonstrate their effect on tumor growth inCRC tumor model. CRC tumor model SW48 xenografts were generated byinjecting 2.5×10⁶ corresponding cultured cells in the presence ofmatrigel subcutaneously into 6-8 weeks old female Ncr-nude mice(Taconic). When most tumors reach 250-300 mm³ in size (˜12 days aftercell inoculation), mice are randomized into treatment groups of 8-10mice using an R studio script designed to group the animals so that theaverage tumor size and standard deviation of the tumor size betweengroups are comparable.

The animals were then injected intravenously with vehicle, human IgG1(isotype control)-MMAE and 4 human CDCP1 ADCs (18C6-MMAE, 27H10-MMAE,38E11-MMAE and 41A9-MMAE) at 5 mg/kg. The i.v. treatment was repeated 3times over a period of 10 days. Following treatment, tumor volumes andmouse weights were monitored until tumors exceed 1500 mm³. Mice thatshowed a 15% or more weight loss or become lethargic are taken out ofthe study.

The 4 ADCs caused complete tumor regression and a significant prolongedsurvival in colon cancer model SW48 tumors (FIG. 11 ). Median of Time toEvent (TTE) of CDCP1 ADC treated groups is 100 days versus 13 and 20days in vehicle and isotype control treated groups, respectively (FIG.12 ).

Example 20. Dose-Dependent Tumor Growth Inhibitory Effects of Anti-CDCP1Antibodies Conjugated to MMAE on SW48 Xenograft Tumors in Mice

Four human CDCP1 antibodies conjugated to MMAE were tested todemonstrate their dose dependent inhibitory effects on tumor growth inSW48 models. The CRC tumor model SW48 xenografts were generated byinjecting 2.5×10⁶ corresponding cultured cells in the presence ofmatrigel subcutaneously into 6-8 weeks old female Ncr-nude mice(Taconic, Hudson, N.Y.). When most tumors reach 250-300 mm³ in size,mice are randomized into treatment groups of 6-8 mice using an R studioscript designed to group the animals so that the average tumor size andstandard deviation of the tumor size between groups arc comparable.

The animals were then injected intravenously with vehicle, human IgG1(isotype control)-MMAE and 4 human CDCP1 ADCs (18C6-MMAE, 27H10-MMAE,38E11-MMAE and 41A9-MMAE) at 0.5, 1.5 and 5 mg/kg. The intravenousadministration was repeated 3 times over a period of 10 days. Followingthe treatment, tumor volumes and mouse weights were monitored untiltumors exceeded 1500 mm³. Mice that showed a 15% or more weight loss orbecome lethargic were taken out of the study. Tumor growth inhibition(TGI) was assessed when the number of animals in the vehicle group(control group) was reduced to <75% and was calculated by the following:TGI=[1−(TV_(tx) _(Day x) −TV_(tx) _(Initial) )/(TV_(veh) _(Day x)−TV_(veh) _(Initial) )]×100%wherein, TV_(tx) _(Initial) is the initial tumor volume in the treatmentgroup, TV_(tx) _(Day x) is the tumor volume in the treatment group atDay X, TV_(veh) _(Initial) is the initial tumor volume in the vehiclegroup and TV_(veh) _(Day x) is the tumor volume in the vehicle group atDay X.

Three out of four human antibody MMAE conjugates, 38E11-MMAE, 27H10-MMAEand 18C6-MMAE, exhibited dose dependent inhibitory effects on SW48 tumorgrowth: at 5 mg/kg, they caused complete tumor regression without tumorregrowth 100 days after initial dosing (FIGS. 13A-13D); at 1.5 mg/kgthese ADCs caused significant tumor growth inhibition: 121%, 106% and117%, respectively, as well as significant prolonged survival, reflectedby the median time to the end point (TTE) as 86, 57 and 51 days,respectively; at 0.5 mg/kg, no obvious tumor growth inhibitory effectwas observed for any of the ADCs. Among the 4 ADCs, 10E2-MMAE was theleast efficacious one with a TGI of 86% and median TTE of 55 days at 5mg/kg (Table 10, FIGS. 13A-13D and 14A-14D).

TABLE 10 Dose dependent tumor growth inhibitory effects of 4 ADCs onestablished SW48 tumors DAR Median TTE (drug:mAb TGI on (time to theTreatment ratio) Day 18 end point) PBS NA NA 16.5 hIgG1-MMAE 5 mg/kg4.16 Unmeasurable 19 38E11-MMAE 0.5 mg/kg 4.25 Unmeasurable 1438E11-MMAE 1.5 mg/kg 4.25 121% 86 38E11-MMAE 5 mg/kg 4.25 125% 10027H10-MMAE 0.5 mg/kg 3.67 Unmeasurable 12.5 27H10-MMAE 1.5 mg/kg 3.67106% 57 27H10-MMAE 5 mg/kg 3.67 123% 100 18C06-MMAE0.5 mg/kg 4.19  50%18 18C6-MMAE 1.5 mg/kg 4.19 117% 51 18C6-MMAE 5 mg/kg 4.19 124% 10010E2-MMAE 0.5 mg/kg 4.16 Unmeasurable 12.5 10E2-MMAE 1.5 mg/kg 4.16Unmeasurable 12.5 10E2-MMAE 5 mg/kg 4.16 86.80%  55

Example 21. Conjugation of Human Antibodies to Pyrrolobenzodiazepine(PBD)

Human monoclonal antibodies against CDCP1 were conjugated toPyrrolobenzodiazepine (PBD) as previously described (Stefano J. E.,Busch M., Hou L., Park A., Gianolio D. A. (2013) Micro- and Mid-ScaleMaleimide-Based Conjugation of Cytotoxic Drugs to Antibody Hinge RegionThiols for Tumor Targeting. In: Ducry L. (eds) Antibody-Drug Conjugates.Methods in Molecular Biology (Methods and Protocols), vol 1045. HumanaPress, Totowa, N.J.) Briefly, each antibody was partially reduced withan appropriate molar amount of TCEP for 2 hrs at 37° C. Afterincubation, the antibodies were cooled to room temperature and a 6-foldmolar excess of PBD was added to the partially reduced antibody for 1hour. After this incubation, the entire mixture was run over a gelfiltration column to remove free drug. A fraction of the eluate was thenused for concentration and DAR determination using A280 and HICrespectively.

Example 22. In Vitro Killing Activity of 38E11 Conjugated to MMAE or PBDon Prostate and Lung Cancer Cells

Human antibodies conjugated to MMAE or PBD as described above are testedfor their effect on growing prostate and non-small cell lung cancercells. Prostate cell lines, DU145, LNCAP, and PC3, and NSCLC cells,which express CDCP1, breast cancer cell line MCF7, which does notexpress CDCP1, were seeded onto 96 well plate at 1000-2000 cells/well.24 hrs later, 38E11-vcMMAE and isotype control hIgG-vcMMAE bearing adrug antibody ratio (DAR) of 3.9 and 4.2, respectively, or 38E11-PBD andisotype control hIgG-PBD bearing a drug antibody ratio (DAR) of 2.3 and2.9, respectively, were added to the wells in complete culture medium sothat the final Ab-PBD concentration in the well is 0.001, 0.01, 0.1 and1 ug/ml. Each treatment was replicated in 2 wells. 96 hrs later, cellviability was measured by CellTiter Glo Luminescent Cell viability assay(Promega) according to manufacturer's instructions. Cell viability wasgraphed by Prism using ratio of cell viability of test conditions tothat of control wells that are treated with growth medium only.

As shown in FIGS. 21A-21B, while 38E11-vcMMAE exhibited no effect onMCF7 cells, which does not express CDCP1, it exhibited killing effectson prostate cancer cell lines, PC3 and DU145, with EC50 of 0.1 μg/ml and1 μg/ml, respectively. LNCAP shows no sensitivity to 38E11-vcMMAE.

As shown in FIGS. 22A-22B, while 38E11-PBD exhibited no effect on MCF7cells, which does not express CDCP1, it exhibited varying degree ofkilling effects on prostate cancer cell lines, PC3, DU145 and LNCAP,with EC50 of 1 μg/ml, 0.1 μg/ml and 0.01 μg/ml, respectively.

As shown in FIGS. 23A-23B, while 38E11-vcMMAE exhibited varying killingeffects on NSCLC cell lines, Cal-12T, HCC78. HCC827, HCC44, HCC15 andH1650 cells, with EC50 more than 1 μg/ml. In contrast, NSCLC cell linesH460 and H1299 showed no sensitivity to 38E11-vcMMAE.

As shown in FIGS. 24A-24B, while 38E11-PBD exhibited varying killingeffects on NSCLC cell lines, with EC50 of ˜0.1 μg/ml in H460 and HCC15cells, and EC50 of 1 μg/ml in HCC15, HCC44 and H1650 cells, and EC50 ofmore than 1 μg/ml inCal-12T, HCC827, H1299.

Example 23. Conjugation of Human Antibodies to Pyrrolobenzodiazepine andAuristatin-E Payloads

Human monoclonal antibodies against CDCP1 were conjugated to thevaline-alanine-pyrrolobenzodiazepine, MA-PEG8-VA-PAB-SG3199 (PBD) ormaleimidocaproyl-valine-citrulline-p-aminobenzoyloxycarbonyl-monomethylauristatin E (MMAE) as previously described (Stefano J. E., Busch M.,Hou L., Park A., Gianolio D. A. (2013) Micro- and Mid-ScaleMaleimide-Based Conjugation of Cytotoxic Drugs to Antibody Hinge RegionThiols for Tumor Targeting. In: Ducry L. (cds) Antibody-Drug Conjugates.Methods in Molecular Biology (Methods and Protocols), vol 1045. HumanaPress, Totowa, N.J.) Briefly, each antibody was partially reduced withan appropriate molar amount of TCEP for 2 hrs at 37° C. Afterincubation, the antibodies were cooled to room temperature and a 6-foldmolar excess of PBD or MMAE was added to the partially reduced antibodyfor 1 hour. After this incubation, the entire mixture was run over a gelfiltration column to remove free drug. A fraction of the eluate was thenused for concentration and DAR determination using A280 and HICrespectively.

Example 24. Dose-Dependent Tumor Growth Inhibitory Effects of 38E11 andhIgG1 Conjugated to MMAE on TNBC PDX Tumors in Mice

Eight monoclonal CDCP1 antibodies conjugated to MMAE and human IgG1 weretested to demonstrate their dose dependent inhibitory effects on tumorgrowth in TNBC PDX models. Xenograft tumors for each PDX model wereinitiated from Cryo-preserved PDX tumor tissue using trocar needlemethod into 3-5 stock mice. Tumor volume was monitored and calculatedusing the formula:Tumor Volume (mm³)=0.52×L×W²where L=length and W=width in mm of a tumor. Tumor length and widthrepresent the two longest perpendicular axes in the x/y plane of eachtumor measured by an electronic caliper to the nearest 0.1 mm.

When tumor volume of stock mice reach 800-1000 mm3, tumors wereharvested for re-implantation into 12-15 pre-study mice. Pre-study miceare implanted unilaterally on the left flank with tumor fragmentsharvested from stock mice. Pre-study tumor volumes are recorded for eachexperiment beginning seven to ten days after implantation. When tumorsreach an average tumor volume of approximately 150-300 mm³ animals arematched by tumor volume into treatment or control groups to be used fordosing.

Human monoclonal antibodies against CDCP1, 38E11, and a human IgG1isotype control antibody (against Hepatitis B virus, HBV) wereconjugated to the anti-mitotic payload,valine-citrulline-monomethylauristatin E (vcMMAE) as previouslydescribed in Stefano, J. E., et al. (2013) Micro- and Mid-ScaleMaleimide-Based Conjugation of Cytotoxic Drugs to Antibody Hinge RegionThiols for Tumor Targeting; and Ducry L. (eds) Antibody-Drug Conjugates.Methods in Molecular Biology (Methods and Protocols), vol. 1045. HumanaPress, Totowa, N.J. Briefly, each antibody was partially reduced with anappropriate molar amount of TCEP for 2 hrs at 37° C. After incubation,the antibodies were cooled to room temperature and a 6-fold molar excessof vcMMAE or PBD was added to the partially reduced antibody for 1 hour.After this incubation, the entire mixture was run over a gel filtrationcolumn to remove free drug. A fraction of the eluate was then used forconcentration and DAR determination using A280 and HIC respectively.Conjugated antibodies arc stored in PBS at 4° C. Dilutions of 1 mg/mlADCs were made in PBS and stored at 4° C. before and during mousetreatment under sterile conditions.

The treatment plan is summarized in the Table 11. For each group,treatments were administered intravenously (i.v.) 2 times per week for 2weeks (2×/wk for 2 wks). Specifically, doses were administered on Day 0,3, 7 and 10. Group 1 received vehicle (PBS), Groups 2 and 3 received 1.5mg/kg or 5 mg/kg of 38E11-vcMMAE, respectively, Group 4 received 5 mg/kgisotype control hIgG1-vcMMAE. Each dose of drug was given in a volume of0.15 ml so that each mouse received indicated doses, assuming a bodyweight of 30 grams for each mouse.

TABLE 11 Treatment Design for TNBC PDX Efficacy Study Total Dose DoseNumber (mg/kg/ Volume of Group -n- Agent dose) (mL/kg) ROA ScheduleDoses 1 2 PBS — 5 IV BIW × 2 4 2 1 38E11- 1.5 5 IV BIW × 2 4 vcMMAE 3 138E11- 5 5 IV BIW × 2 4 vcMMAE 4 1 hIgG- 5 5 IV BIW × 2 4 vcMMAE

Eight TNBC PDX models (CTG-1017, CTG-1520, CTG-1646, CTG-0888, CTG-0437,CTG-1374 and CTG-1883) were dosed and analyzed. The response of each ofthe eight models to different treatment are summarized in Table 12.FIGS. 25A-25B shows the tumor growth curves of these 8 models. FIGS.26A-26B shows the body weight change of mice from these eight modelsthroughout the study.

TABLE 12 Response Summary of MDA-MB-231 Xenograft Efficacy Study % tumorsize vs vehicle (positive no.) or % regression (negative no.) 38E11-38E11- hIgG- Tumor Target vcMMAE vcMMAE vcMMAE Model ID Statusexpression 1.5 mg/kg 5 mg/kg 5 mg/kg CTG-1017 Primary 1+ 31% −29% −35%CTG1520 Relapsed 1+  0% −52% −38% CTG-1646 Primary 1+ −86%  −83%  59%CTG-0888 Relapsed 2+ 102%  −10% −46% CTG-2055 Relapsed 2+ 51%  70%  92%CTG-0437 Mets 3+ −87%  −87% −24% CTG-1374 Primary 3+ 45% −60%  −7%CTG-1883 Primary 3+ 17%  29%  54%Response of TNBC PDX models to 38E11-vcMMAE and hIgG-vcMMAE

Seven out of eight TNBC PDX models (CTG-1017, CTG-1520, CTG-1646,CTG-0888, CTG-0437, CTG-1374 and CTG-1883) responded to 38E11-vcMMAE at5 mg/kg. These models showed response of ΔT/ΔC<20% or regression ordisease control as shown in Table 12 and FIGS. 25A-25B. Four out ofeight models (CTG-1520, CTG-1646, CTG-0437, and CTG-1883) responded to38E11-vcMMAE at 1.5 mg/kg. These models showed response of ΔT/ΔC<20% orregression or disease control as shown in Table 12 and FIGS. 25A-25B.Seven out of eight TNBC PDX models (CTG-1017, CTG-1520, CTG-0888,CTG-0437, and CTG-1374) responded to hIgG-vcMMAE at 5 mg/kg. Thesemodels showed response of ΔT/ΔC<20% or regression or disease control asshown in Table 12 and FIGS. 25A-25B.

INCORPORATION BY REFERENCE

The contents of all references, patents, pending patent applications andpublished patents, and Accession Numbers, cited throughout thisapplication are hereby expressly incorporated by reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

The invention claimed is:
 1. An antibody or antigen-binding fragmentthereof that binds to CUB domain-containing protein 1 (CDCP1), whereinthe antibody or antigen-binding fragment thereof comprises a heavy chainvariable region comprising HCDR1, HCDR2, and HCDR3, and a light chainvariable region comprising LCDR1, LCDR2, and LCDR3, wherein HCDR1,HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ IDNOs: 155, 156, and 157 respectively, and LCDR1, LCDR2, and LCDR3comprise the amino acid sequences set forth in SEQ ID NOs: 158, 159, and160, respectively.
 2. An antibody or antigen-binding fragment thereofthat binds to CUB domain-containing protein 1 (CDCP1), wherein theantibody or antigen-binding fragment thereof comprises a heavy chainvariable region (VH) and a light chain variable region (VL), wherein theVH comprises an amino acid sequence that is at least 90% identical toSEQ ID NO: 151, wherein the VH comprises HCDR1, HCDR2, and HCDR3comprising the amino acid sequences set forth in SEQ ID NOs: 155, 156,and 157, respectively, and the VL sequence comprises an amino acidsequence that is at least 90% identical to SEQ ID NO: 152, wherein theVL comprises LCDR1, LCDR2, and LCDR3 comprising the amino acid sequencesset forth in SEQ ID NOs: 158, 159, and 160, respectively.
 3. Theantibody or antigen-binding fragment thereof of claim 2, wherein the VHcomprises the sequence of SEQ ID NO: 151 and the VL comprises thesequence of SEQ ID NO:
 152. 4. A pharmaceutical composition comprisingan antibody or antigen-binding fragment thereof of claim 1, and apharmaceutically acceptable carrier.
 5. An isolated nucleic acidencoding the antibody or antigen-binding fragment thereof of claim
 1. 6.A vector comprising the nucleic acid of claim
 5. 7. A host cellcomprising the vector of claim
 6. 8. A method of producing an antibodyor antigen-binding fragment thereof, the method comprising: culturingthe host cell of claim 7 under conditions wherein said antibody orantigen-binding fragment thereof is expressed; and recovering saidantibody or antigen-binding fragment thereof.
 9. A pharmaceuticalcomposition comprising an antibody or antigen-binding fragment thereofof claim 2, and a pharmaceutically acceptable carrier.
 10. An isolatednucleic acid encoding the antibody or antigen-binding fragment thereofof claim
 2. 11. A vector comprising the nucleic acid of claim
 10. 12. Ahost cell comprising the vector of claim
 11. 13. A method of producingan antibody or antigen-binding fragment thereof, the method comprising:culturing the host cell of claim 12 under conditions wherein saidantibody or antigen-binding fragment thereof is expressed; andrecovering said antibody or antigen-binding fragment thereof.
 14. Apharmaceutical composition comprising an antibody or antigen-bindingfragment thereof of claim 3, and a pharmaceutically acceptable carrier.15. An isolated nucleic acid encoding the antibody or antigen-bindingfragment thereof of claim
 3. 16. A vector comprising the nucleic acid ofclaim
 15. 17. A host cell comprising the vector of claim
 16. 18. Amethod of producing an antibody or antigen-binding fragment thereof, themethod comprising: culturing the host cell of claim 17 under conditionswherein said antibody or antigen-binding fragment thereof is expressed;and recovering said antibody or antigen-binding fragment thereof. 19.The antibody or antigen-binding fragment thereof of claim 1, wherein theantibody or antigen-binding fragment thereof is an IgG1 isotype.